СРЕДА ДЛЯ КУЛЬТИВИРОВАНИЯ КЛЕТОК БЕЗ БЕЛКОВ И БЕЗ СЫВОРОТКИ

Изобретение относится к биотехнологии и может быть использовано при культивировании клеток млекопитающих, а также для получения белка из клеток млекопитающих. Питательная среда представляет собой синтетическую среду с добавлением гидролизата сои в количестве от 0,1 до 100 г/л, причем по меньшей мере 40% гидролизата имеет м.м. ≤500 Да. Среда может также содержать буфер, стабилизатор окисления и т.д., а синтетической средой является среда DMEM/HAM F12, 199 или RPMI. При выращивании клеток в данной среде увеличивается продукция как рекомбинантных клеток, так и их продуктивность (увеличение выхода белка). Кроме того, среда является универсальной в плане выбора метода культивирования клеток млекопитающего. 3 н. и 11 з.п. ф-лы, 5 ил., 7 табл.

СПОСОБ ПОЛУЧЕНИЯ АНТИТЕЛА ИЛИ ЕГО ФРАГМЕНТА С ПОДПИТКОЙ (ВАРИАНТЫ)

Изобретение относится к области получения рекомбинантных белков и касается способов их получения. Представлены способы получения антител, включающие: культивирование клеток млекопитающего, содержащих нуклеиновую кислоту, кодирующую антитело или его фрагмент, в культуре клеток, содержащей продукционную среду культуры клеток; подпитку клеток млекопитающего добавлением обогащенного гидролизатом раствора и обогащенного раствора базальной среды к культуре клеток, где обогащенный гидролизатом раствор содержит гидролизат продуктов растительного происхождения и 75-300 г/мл гидролизата дрожжей. Представленными способами возможно получать, в частности, такие антитела, как анти-TNFα-антитело или анти-IL-12-антитело. Охарактеризованное решение позволяет получить повышенное количество антител и может быть использовано в фармацевтической промышленности. 3 н. и 25 з.п. ф-лы, 46 табл., 4 пр., 1 ил.

MYELOMA CELL CULTURE IN A TRANSFERRIN FREE MEDIUM WITH LOW EISENGEHALT

BASAL MEDIUM FOR THE CULTIVATION OF IT CELLS

CHEMICALLY DEFINED MEDIUM FOR CULTIVATED MAMMALIAN CELLS

CELL CULTURE WITH NM23, NM23 COMPREHENSIVE ONE CELL CULTURAL MEDIA AS WELL AS THERAPEUTIC USE OF CELLS CULTIVATED IN PRESENCE OF NM23

NICHTTUMORBILDENDE MDCK ZELLINIE ONES FOR THE INCREASE OF VIRUSES

SERUM-FREE MEDIUM

Method of increasing the expression yield of vitamin K-dependent proteins

The invention encompasses the use of one or more compounds selected from a list comprising i) reduced forms of vitamin K and/or ii) reduced forms of a vitamin K analog and/or iii) reduced forms of a vitamin K precursor for the expression of one or more functional vitamin K-dependent proteins in cell culture as well as processes for the fermentation of eucaryotic cells expressing one or more vitamin K-dependent proteins wherein one or more compounds selected from a list comprising i) reduced forms of vitamin K and/or ii) reduced forms of a vitamin K analog and/or iii) reduced forms of a vitamin K precursor are added to the cell culture medium before and/or during the fermentation process.

Methods of generating T-cells from stem cells and immunotherapeutic methods using the T-cells

Methods and composition for production of T cells are provided. Also provided are therapeutic methods using engineered T cells. For example, in certain aspects methods include preparing three dimensional cell culture compositions comprising stroma cells and hematopoietic stem or progenitor cells in a serum-free medium for producing T cells.

CELL CULTURE MEDIUM FOR ENHANCED CELL GROWTH, CULTURE LONGEVITY AND PRODUCT EXPRESSION

COLOSTRUM FRACTION, PREPARATION AND USE THEREOF AS CELL CULTURE MEDIA SUPPLEMENT

PERFUSION MEDIUM

The invention relates to a method of culturing mammalian cells expressing a heterologous protein in a perfusion cell culture comprising increasing the potassium concentration and decreasing the molar ratio of sodium to potassium to reduce wasteful cell bleed and to increase protein production. The invention further relates to a serum-free perfusion medium comprising a high potassium ion concentration and a low molar ratio of sodium to potassium and to the use of this medium for use in culturing cells in a perfusion culture during production phase or for reducing the cell bleed volume during production phase.

METHOD FOR DETECTING OR SEPARATING/OBTAINING CIRCULATING TUMOR CELL EMPLOYING CELL PROLIFERATION METHOD

The present invention addresses the problem of providing a method for detecting or separating/obtaining a circulating tumor cell (CTC), whereby it becomes possible to reliably and steadily detect or separate/obtain a CTC and a circulating tumor stem cell (CTSC) which are present in trace amounts in a biological circulating body fluid, such as blood and lymph fluid, even under the condition where the type of the tumor cell is not identified yet and the tumor cell is present in a trace amount in the biological circulating body fluid. The problem can be solved by a method for detecting or separating/obtaining a CTC and/or a CTSC in a biological circulating body fluid, said method comprising the following treatment steps (1) to (4): (1) a first step of pretreating a sample collected from the biological circulating body fluid to obtain a mononuclear cell phase; (2) a second step of providing a well plate that is prepared by injecting a liquid culture medium comprising a serum-free culture medium ...

EXTRACT OF UNDIFFERENTIATED CELLS OF MIMOSA PUDICA AND USES THEREOF IN DERMO-COSMETICS

La présente invention concerne une préparation issue d'une culture in vitro de cellules indifférenciées de Mimosa pudica ainsi que son procédé de préparation; une composition cosmétique ou dermatologique comprenant ladite préparation; et ses utilisations pour le traitement des troubles inflammatoires cutanés, comme agent anti-oxydant dont le traitement des stress oxydatifs dus à la pollution environnementale, et comme agent anti-âge ...

SEED TRAIN PROCESSES AND USES THEREOF

Provided herein are seed train processes and methods of producing a recombinant protein that include the use of these seed train processes.

MAMMALIAN CELL CULTURE MEDIA WHICH COMPRISE SUPERNATANT FROM COHN FRACTIONATION STAGES AND USE THEREOF

... ²²The present invention relates to mammalian cell culture media which comprise ²supernatant ²from some of the fractions of human plasma fractionation according to the Cohn ²method, ²more specifically, the supernatant of fractions I and II+III. When said ²supernatant is added ²as a culture medium supplement it provides various nutrients and factors for ²the effective ²maintenance and/or proliferation of the cultured mammalian cells. In addition, ²the present ²invention relates to the preparation process and use of said medium in the ²culture of ²mammalian cells.² ...

METHOD FOR PRODUCING ADENOHYPOPHYSIS OR PRECURSOR TISSUE THEREOF

The present invention provides a method for deriving an adenohypophysis or precursor tissue thereof in vitro from human pluripotent stem cells. In this method: a human cell aggregate including hypothalamus neuroepithelial tissue and epidermal ectoderm is obtained by subjecting a human pluripotent stem cell aggregate to suspension culture in a culture medium including a bone morphogenetic factor signal transduction pathway activator and an Shh signal pathway active substance; and by subjecting the obtained human cell aggregate including hypothalamus neuroepithelial tissue and epidermal ectoderm to further suspension culture in a culture medium including a bone morphogenetic factor signal transduction pathway activator and an Shh signal pathway active substance, the formation of a hypophysis placode and/or Rathke's pouch in the epidermal ectoderm is induced to obtain a human cell aggregate including 1) hypothalamus neuroepithelial tissue and 2) a hypophysis placode and/or Rathke's pouch.

IMPROVED CULTURE MEDIA ADDITIVE AND PROCESS FOR USING IT

The present invention relates to an improved cell culture additive with a content of polyamines and iron, media containing it and processes for using it for an improved cell growth, cell viability or cellular productivity.

CELL CULTURE IMPROVEMENTS

... ²²²The invention describes improved methods and compositions for producing a ²recombinant ²protein, e.g., an antibody, in mammalian cell culture. In addition, the ²invention provides ²improved cell culture media, including improved production media, feed ²solutions, and ²combination feeds, which may be used to improve protein productivity in ²mammalian cell ²culture.² ...

METHOD OF ENHANCING CELL GROWTH USING ALKYL-AMINE-N-OXIDE (AANOX)

The present invention relates to a method to enhance cell growth in culture comprising adding an alkyl-amine-n-oxide (AANOx), such as dodecyldimethylamine oxide (DDAO), into the culture medium in an amount sufficient to improve cell growth.

METHODS FOR ENHANCED PRODUCTION OF BONE MORPHOGENETIC PROTEINS

Methods and processes for improved recombinant protein production are provided. The methods are useful for production of growth factors, particularly those of the TGF-.beta. superfamily, including bone morphogenetic proteins (BMPs), such as BMP-2. Suitable host cells are cultured in media where iron is present at a concentration of at least 2.25 µM and if pyridoxal is present, it makes up less than 55 % of the molar concentration of vitamin B6 in the media.

CULTIVATION OF CELLS OF MAMMALS

MEDIUM FOR CULTIVATION OF CELLS

METHODS OF REGULATING THE CONTENT OF MANNOSE BEFORE THE RECOMBINANT PROTEINS

Method for producing engineered heart muscle (EHM)

The vitamin K-dependent protein of the output method of the expression

Establishment for serum-free freezing medium and adipose-derived stem cell library of adipose-derived stem cells

Improved culture media additive and process for using it

ORGAN PRESERVATIVE COMPOSITION AND USES THEREOF

AGENT ACTIVATOR OF THE SPECIFIC PRODUCTIVITY OF THE ANIMAL CELLS RECOMBINING CONTAINING POLYVINYLPYRROLIDONE AND DEFINITE CULTURE MEDIUM THE CONTAINER

배양 배지에서 식물 단백질 상동체의 활용

... 본 개시내용은 부분적으로, 혈청 단백질의 적어도 하나의 식물 단백질 상동체를 포함하는 세포 배양 배지 보충물, 무혈청 기본 배지 및 하나 이상의 식물 기반 단백질을 포함하는 세포 배양 배지, 및 시험관 내에서 세포를 성장시키고 세포 배양 배지를 사용하여 배양육을 생산하는 방법을 제공한다.

MEDIOS ENRIQUECIDOS CON NUTRIENTES PARA EL CULTIVO DE HUTC

Ésta proporciona métodos para cultivar células dependientes del anclaje (por ejemplo, hUTC) en medio de cultivo que comprende aminoácidos, vitaminas, sales, nucleósidos, insulina, transferrina, etanolamina y selenio de sodio, en donde el medio de cultivo se complementa con suero. El método comprende, además, la adición de una solución nutriente libre de suero que comprende aminoácidos, vitaminas, sales, nucleósidos, insulina, transferrina, etanolamina y selenio de sodio. La presente proporciona, además, medios de cultivo y soluciones nutrientes libres de suero para cultivar células dependientes del anclaje. Reivindicación 1: Un medio de cultivo para el crecimiento de células dependientes del anclaje caracterizado porque comprende: los aminoácidos L-arginina; L-cistina; L-cisteína; L-glutamina; glicina; L-histidina; L-isoleucina; L-leucina; L-lisina; L-metionina; L-fenilalanina; L-serina; L-treonina; L-triptófano; L-tirosina; L-valina; L-alanina; L-esparagina; ácido L-aspártico; ácido L-glutámico ...

PROCESS FOR THE PREPARATION OF GLYCOSYLATED INTERFERON BETA

Serum-free culture medium for the production of recombinant gonadotropins

The present invention is in the field of the manufacture of recombinant proteins. More specifically, it relates to the use of a serum-free culture medium comprising an antioxidant for the production of recombinant gonadotropins. The antioxidant may be selected from the group consisting of L-glutathione, 2-mercaptoethanol, L-methionine and a combination of ascorbic acid and of (+)-alpha-tocopherol.

METHODS FOR ENHANCED PRODUCTION OF BONE MORPHOGENETIC PROTEINS

PHARMACEUTICAL COMPOSITION FOR TREATING ISCHEMIC DISEASES, CONTAINING CONDITIONED MEDIUM OBTAINED THROUGH THREE-DIMENSIONAL CELL CULTURE AS ACTIVE INGREDIENT

The present invention provides a pharmaceutical composition for treating ischemic diseases, containing a conditioned medium obtained from a cultured product obtained through three-dimensional culturing of adult stem cells in a medium for cell culture, as an active ingredient. The concentration and content of an angiogenesis promoting factor of the conditioned medium according to the present invention is very high compared with those of a conditioned medium obtained through a two-dimensional culture method, and thus the conditioned medium of the present invention can be useful for the vascular regeneration in ischemic vascular diseases. In addition, the conditioned medium can be clinically applied safely since there are no animal-derived ingredients, a buffer solution and indicator ingredients which have been used in a conventional cell culture medium or conditioned medium, and the conditioned medium can be effectively used in ischemic vascular diseases and the like even in small amounts ...

Production of recombinant factor VIII in the presence of liposome-like substances of mixed composition

Recombinant Factor VIII expression in a mammalian cell culture can be increased by including a novel liposome-like substance in the culture medium. The liposome-like substance comprises at least two (preferably at least three) different lipids in defined molar ratios. In a preferred embodiment, the addition of a liposome-like substance comprised of dioleoyl phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine in a molar ratio of 4:1:1 to the culture medium of GS-MDR cells resulted in an increase in FVIII production by a factor greater than five.

Methods of expanding embryonic stem cells in a suspension culture

A method of expanding and maintaining human embryonic stem cells (ESCs) in an undifferentiated state by culturing the ESCs in a suspension culture under culturing conditions devoid of substrate adherence is provided. Also provided are a method of deriving ESC lines in the suspension culture and methods of generating lineage-specific cells from ESCs which were expanded in the suspension culture of the present invention.

COMPOSITIONS COMPRISING SMALL EXTRACELLULAR VESICLES DERIVED FROM UMBILICAL CORD BLOOD MONONUCLEAR CELLS WITH ANTI-INFLAMMATORY AND IMMUNOMODULATORY PROPERTIES AND PROCESS FOR OBTAINING THEM

The present invention relates to a process for isolating Small Extracellular Vesicles secreted by umbilical cord blood mononuclear cells (UCBMNCs) and compositions comprising said Small Extracellular Vesicles, which are useful to be applied to autoimmune diseases therapeutics or prophylactics and/or cosmetic purposes. The proposed process for isolating UCBMNCs Small Extracellular Vesicles comprises three main steps: i) a first step of sequential centrifugation, ii) a second step of microfiltration combined with ultrafiltration (UF), and iii) a third step of size exclusion chromatography (SEC) and aims to achieve highly pure Small Extracellular Vesicles and in a higher yield. The SEVs compositions comprise specific type of proteins, RNA and lipids, that enables them to be very effective when applied to inflammatory and autoimmune diseases therapeutics, such as psoriasis, lupus, atopic dermatitis, eczema, etc. and also to cosmetic or prophylactic compositions. Therefore, the present invention ...

Media for Cell Culture

The present disclosure relates, in general, to a media, e.g., a serum replacement, media supplement, complete media or cryopreservation media, comprising a base physiological buffer and liposomes comprising cholesterol, phosphatidylcholine and fatty acids. It is contemplated that media provides advantages to improve cell growth in culture compared to cells cultured not using the serum replacement described herein. 1. A media for use with cells in suspension or in adherent culture , the media comprising a base physiological buffer liquid mix and(a) liposomes comprising cholesterol, phosphatidylcholine and fatty acids, wherein the liposome is in an amount such that the final concentration of cholesterol in a cell suspension or adherent culture is from 1 to 20 mg/L, and wherein the final concentration of phosphatidylcholine in a cell suspension or adherent culture is from 100 to 1000 mg/L; or(b) pectin; or(a) and (b).2. The media of claim 1 , wherein the liposome comprises one or more fatty acids selected from the group consisting of linolenic acid claim 1 , linoleic acid claim 1 , myristic acid and oleic acid.3. The media of claim 1 , wherein the liposome further comprises ethanolamine and polysorbate.4. The media of claim 1 , wherein the final concentration of pectin in a cell suspension or adherent culture is from 25 to 500 mg/L.5. The media of claim 1 , wherein the base physiological buffer liquid mix comprises one or more organic salt claim 1 , inorganic salt claim 1 , buffer claim 1 , iron source or iron transporter claim 1 , glycerol claim 1 , amino acid claim 1 , vitamin claim 1 , sugar claim 1 , antioxidant and trace element.612-. (canceled)13. The media of claim 1 , wherein the media is a serum replacement claim 1 , complete media claim 1 , media supplement or cryopreservation media.14. A serum replacement claim 1 , complete media or media supplement comprising liposomes and a base physiological buffer liquid mix claim 1 , wherein the liposomes comprise ...

Use of esters of unsaturated, physiologically active fatty acids as nutrient media for cell cultures

The disclosed invention relates to the use of esters of unsaturated, physiologically active fatty acids as nutrient media for cell cultivation and, more particularly, as a substitute for foetal bovine serum. In one aspect, the esters comprise more than 50 mol-% of physiologically active fatty acids containing 16 to 24 carbon atoms and 2 to 5 double bonds as the acid component and a lower C_1-4 alcohol, preferably ethanol, or a sterol as ester component. In another aspect, the esters comprise a transesterification product of natural or synthetic oils or a mixture of such oils having greater than 50 mol-% of unsaturated, physiologically active fatty acids, based on the acyl group, and a lower C_1-4 alcohol or a sterol. In a further aspect, the esters are used together with sterols, phospholipids and/or vegetable proteins or are liposomally encapsulated.

СПОСОБ КУЛЬТИВИРОВАНИЯ В СИСТЕМЕ ПОСЕВНЫХ ФЕРМЕНТЕРОВ (ВАРИАНТЫ)

Группа изобретений относится к области биотехнологии. Предложен способ получения рекомбинантного белка и его выделения (варианты). Способ включает помещение рекомбинантных клеток млекопитающих в первую культуральную среду для получения первой культуры клеток, периодическое культивирование, помещение первой культуры клеток в перфузионный биореактор со второй культуральной средой для получения второй культуры клеток, перфузионное культивирование, помещение второй культуры клеток в производственный биореактор с третьей культуральной средой для получения культуры клеток-продуцентов и их перфузионное культивирование в позволяющих рекомбинантным клеткам секретировать рекомбинантный белок условиях. В одном варианте способа используют содержащий рекомбинантные клетки замороженный клеточный банк. При этом рекомбинантный белок выделяют из третьей культуральной среды, удаленной из производственного биореактора. Изобретения обеспечивают снижение числа стадий культивирования, количества времени от начальной ...

СПОСОБ ОБНАРУЖЕНИЯ ИЛИ ВЫДЕЛЕНИЯ/ПОЛУЧЕНИЯ ЦИРКУЛИРУЮЩЕЙ ОПУХОЛЕВОЙ КЛЕТКИ, ИСПОЛЬЗУЮЩИЙ МЕТОД ПРОЛИФЕРАЦИИ КЛЕТОК

Изобретение относится к медицине, а именно к способу обнаружения циркулирующей опухолевой клетки и/или циркулирующей опухолевой стволовой клетки из биологической циркулирующей жидкости организма. Способ включает этап предварительной обработки образца биологической циркулирующей жидкости организма для получения фазы мононуклеарных клеток, в котором предварительная обработка образца биологической циркулирующей жидкости организма с удалением жидкого компонента и неклеточного компонента, содержащегося в биологической циркулирующей жидкости организма; этап получения луночного планшета, в который введена культуральная среда, содержащая бессывороточную ростовую среду для циркулирующей опухолевой клетки и/или циркулирующей опухолевой стволовой клетки, и посева в него мононуклеарных клеток, полученных на первом этапе, с последующей инкубацией; этап удаления культуральной среды из лунки планшета, полученного путем инкубации на втором этапе; а также этап обнаружения адгезивной опухолевой клетки, прикрепленной ...

СРЕДЫ ДЛЯ КУЛЬТУР КЛЕТОК МЛЕКОПИТАЮЩИХ, КОТОРЫЕ СОДЕРЖАТ НАДОСАДОЧНУЮ ЖИДКОСТЬ СТАДИЙ ФРАКЦИОНИРОВАНИЯ ПО КОНУ, И ИХ ПРИМЕНЕНИЕ

... 1. Среда для культивирования клеток млекопитающих, характеризующаяся тем, что, в дополнение к обычным питательным веществам в основной культуральной среде для культивирования клеток млекопитающих, она содержит надосадочную жидкость одной из стадий фракционирования плазмы человека по методу Кона. ! 2. Культуральная среда по п.1, характеризующаяся тем, что надосадочная жидкость представляет собой надосадочную жидкость фракции I по методу Кона. ! 3. Культуральная среда по п.1, характеризующаяся тем, что надосадочная жидкость представляет собой надосадочную жидкость фракции II+III по методу Кона. !4. Культуральная среда по любому из предшествующих пунктов, характеризующаяся тем, что надосадочные жидкости фракций по Кону получают из смесей (пулов) от по меньшей мере 1000 людей-доноров. ! 5. Культуральная среда по п.1, характеризующаяся тем, что надосадочную жидкость фракции по Кону высушивают. ! 6. Культуральная среда по по п.1, характеризующаяся тем, что надосадочную жидкость фракции по Кону ...

СРЕДА ДЛЯ КУЛЬТИВИРОВАНИЯ КЛЕТОК БЕЗ БЕЛКОВ И БЕЗ СЫВОРОТКИ

... 1. Способ получения рекомбинантного фактора VIII из клеточной культуры, включающий ! введение клеток млекопитающих, которые содержат последовательности, кодирующие рекомбинантный фактор VIII в среду, не содержащую белков и не содержащую сыворотки, содержающую ультрафильтрованный гидролизат сои, где по меньшей мере 40 % указанного гидролизата сои имеет молекулярную массу ≤500 Дальтон, где указанные клетки экспрессируют указанный рекомбинантный фактор VIII; ! выращивание указанных клеток в этой среде и экспрессирование указанного рекомбинантного фактора VIII с получением смеси указанных клеток и указанного рекомбинантного фактора VIII в указанной среде, ! выделение указанного рекомбинантного фактора VIII из этой смеси. ! 2. Способ по п.1, где указанная среда содержит гидролизат сои в количестве превышающем 10 % по весу в пересчете на общий сухой вес среды. ! 3. Способ по п.1, где указанная среда содержит очищенный гидролизат сои. ! 4. Способ по п.1, где гидролизат сои имеет содержание эндотоксина ...

Serumfreies Medium für Säugetierzellen

COLOSTRUM PARLIAMENTARY GROUP AND YOUR USE AS ADDITIVE WITH THE CELL CULTURE MEDIUM

PRODUCTION OF REKOMBINANTEM FACTOR VIII IN PRESENCE OF LIPOSOM SIMILAR CONNECTING COMPOSITIONS

USE FROM NANO-EMULSIONS TO THE DETERMINATION OF THE BIOCOMPATIBILITY OF LIPOPHILIC MATERIALS IN THE CELL CULTURAL TEST AND FOR IT APTITUDE NANO-EMULSIONS

FULLSYNTHETIC CELL CULTURE MEDIUM.

CHOLESTEROLREICHE PARLIAMENTARY GROUP, PROCEDURE FOR IT AND USE THE SAME TO THE CELL CULTURE.

SERUM-FREE MEDIUM FOR MAMMALIAN CELLS

Cell culture method and utilization of the same

It is intended to make cells to produce a protein at a high level by using a medium containing an enzymatically digested product of fish meant or a fish meat extract. Namely, a method of culturing cells characterized by comprising initiating the culture of the cells in a medium containing an enzymatically digested product of fish meant or a fish meat extract, and then adding the enzymatically digested product of fish meant or the fish meat extract to the medium at least once during the cell culture; and a method of producing a desired protein by using this culture method.

METHODS OF PREPARING FEEDER CELLS-FREE, XENO-FREE HUMAN EMBRYONIC STEM CELLS AND STEM CELL CULTURES PREPARED USING SAME

Production of recombinant factor VIII in the presence of lipsome-like substances of mixed composition

Method of enhancing cell growth using alkyl-amine-n-oxide (AANOx)

The present invention relates to a method to enhance cell growth in culture comprising adding an alkyl-amine-n-oxide (AANOx), such as dodecyldimethylamine oxide (DDAO), into the culture medium in an amount sufficient to improve cell growth.

Methods of in vitro oocyte development

Methods of preparing ovarian tissue for primordial follicle growth are presented comprising the steps: providing an ovarian tissue sample comprising cortical tissue and stromal tissue; removing damaged tissue from the ovarian tissue sample where present; removing excess stromal tissue from the ovarian tissue sample where present; and then mechanically stretching the ovarian tissue sample along at least one dimension of the ovarian tissue sample, such that the size of the ovarian tissue sample along the at least one dimension is increased by at least 10%. Methods of growing viable oocyte in vitro, and methods of preparing individual ovarian follicles for growth are also presented.

Methods for modulating production profiles of recombinant proteins

The present invention relates to methods and compositions for modulating glycosylation of recombinant proteins expressed by mammalian host cells during the cell culture process. Also 5 disclosed are methods of culturing a host cell expressing a recombinant protein in a cell culture medium comprising a disaccharide or a trisaccharide, while keeping the osmolality constant.

A CELL CULTURE MEDIUM AND METHODS FOR ENHANCING RECOMBINANT ANTIBODY PURITY

This invention relates to a novel cell culture medium and methods to enhance recombinant antibody purity using the cell culture medium disclosed herein. The novel cell 15 culture medium is aself-made feeding medium, which comprises from about 90 nM to about 500 mM cysteine, from about 50 mM to about 500 mM tyrosine, and from about 50 mM to about 300 mM tryptophan. This invention also relates to a method of growing cell culture using the cell culture medium disclosed herein By controlling the concentration of cysteine in the self-made feed medium as well as the amount and time of adding this feed medium into 0 the cell culture, the purity of antibodies is significantly improved while glycosylation profile and antibody expression level are consistently maintained to guarantee the efficacy of antibodies. '5 -17- 3. Ox1O - 140mM Cys - 210mM Cys 2. 5x107 -A-- 280mM Cys 2. Ox1O 1. 5x10 1. Ox1O 5. Ox1O 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Culture Days (d) Figure 1 - 140mM Cys 210mM Cys 40 - 280mM ...

Method for culturing MDCK cells

The present invention pertains to: cloned MDCK cells which exhibit an expansion factor of 4.5-fold or more when being cultured using microcarriers; a method for culturing the MDCK cells; and a method for multiplying a virus by using the MDCK cell culturing method.

Method for detecting or separating/obtaining circulating tumor cell employing cell proliferation method

The present invention addresses the problem of providing a method for detecting or separating/obtaining a circulating tumor cell (CTC), whereby it becomes possible to reliably and steadily detect or separate/obtain a CTC and a circulating tumor stem cell (CTSC) which are present in trace amounts in a biological circulating body fluid, such as blood and lymph fluid, even under the condition where the type of the tumor cell is not identified yet and the tumor cell is present in a trace amount in the biological circulating body fluid. The problem can be solved by a method for detecting or separating/obtaining a CTC and/or a CTSC in a biological circulating body fluid, said method comprising the following treatment steps (1) to (4): (1) a first step of pretreating a sample collected from the biological circulating body fluid to obtain a mononuclear cell phase; (2) a second step of providing a well plate that is prepared by injecting a liquid culture medium comprising a serum-free culture medium ...

Seed train processes and uses thereof

Abstract Provided herein are seed train processes and methods of producing a recombinant protein that include the use of these seed train processes.

Nutrient enriched media for hUTC growth

This invention provides for methods of growing anchorage-dependent cells (e.g. hUTC) in culture medium comprising amino acids, vitamins, salts nucleosides, insulin, transferrin, ethanolamine and sodium selenium, wherein the culture medium is supplemented with serum. The method further comprises addition of a serum-free nutrient solution comprising amino acids, vitamins, salts nucleosides, insulin, transferrin, ethanolamine and sodium selenium. The invention also provides for culture media and serum-free nutrient solutions for growing anchorage-dependent cells.

METHODS OF PREPARING FEEDER CELLS-FREE, XENO-FREE HUMAN EMBRYONIC STEM CELLS AND STEM CELL CULTURES PREPARED USING SAME

LIPID MICROEMULSIONS FOR CULTURE MEDIA

Lipid microemulsions which can be added to cell culture media to provide essential lipids in a bioavailable form and their components are disclosed. Methods to disperse lipids in culture media by the use of one or more emulsifiers are described. Further disclosed are media which support growth of cells and production of recombinant, viral and/or native products wherein lipids are supplied in the form of a microemulsion.

VERY LOW PROTEIN NUTRIENT MEDIUM FOR CELL CULTURE

This nutrient medium is very effective for the serum-free or serum-protein-free culture of various animal cells, in both high and low density culture. Serum proteins have been replaced with non-protein-based cell growth enhancers and a non-serum derived protein supplement. The non-protein growth enhancer is a modified or derivatized polyurethane prepolymer or polymer and preferably is a sulfhydryl derivative of polyurethane. The protein supplement may be insulin, an insulin analog or an insulin-like growth factor.

SERUM-FREE CELL CULTURE MEDIUM

The specification describes an improved serum-free animal cell culture medium, which can used for the production of a protein of interest. Ornithine, or a combination of ornithine and putrescine can be added to serum-free media or chemically defined media to improve viable cell density, to reduce cell doubling time, and to increase the production of a protein of interest.

MAMMALIAN CELL CULTURE

The invention provides a method for culturing mammalian cells. The method provides greater control over cell growth to achieve high product titer cell cultures by changing the temperature of the cell culture and/or by starving the cells in their asparagine supply.

METHODS FOR MODULATING PRODUCTION PROFILES OF RECOMBINANT PROTEINS

The present invention relates to methods and compositions for modulating glycosylation of recombinant proteins expressed by mammalian host cells during the cell culture process. Also 5 disclosed are methods of culturing a host cell expressing a recombinant protein in a cell culture medium comprising a disaccharide or a trisaccharide, while keeping the osmolality constant.

METHODS FOR MODULATING MANNOSE CONTENT OF RECOMBINANT PROTEINS

The present invention relates to methods of modulating (e.g., reducing) the mannose content, particularly high-mannose content of recombinant glycoproteins.

IMPROVED CULTURE MEDIA ADDITIVE AND PROCESS FOR USING IT

The present invention relates to an improved cell culture additive with a content of polyamines and iron, media containing it and processes for using it for an improved cell growth, cell viability or cellular productivity.

NATIVE WHARTON'S JELLY STEM CELLS AND THEIR PURIFICATION

Noncultured Wharton's Jelly stem cells and methods of their purification, storage and use are provided.

PRODUCTION OF RECOMBINANT FACTOR VIII IN THE PRESENCE OF LIPOSOME-LIKE SUBSTANCES OF MIXED COMPOSITION

Recombinant Factor VIII expression in a mammalian cell culture can be increased by including a novel liposome-like substance in the culture medium. The liposome-like substance comprises at least two (preferably at least three) different lipids in defined molar ratios. In a preferred embodiment, the addition of a liposome-like substance comprised of dioleoyl phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine in a molar ratio of 4:1 :1 to the culture medium of GS-MDR cells resulted in an increase in FVIII production b y a factor greater than five.

SERUM-FREE CELL CULTURE MEDIUM

BEHRINGWERKE AKTIENGESELLSCHAFT 92/B 015 - Ma 930 Auslandstext of the disclosure Serum-free cell culture medium The invention relates to a nutrient medium for growing cells, which contains amino acids, vitamins, cofactors and inorganic salts but is free of animal proteins.

Colostrum Fraction, Preparation and Use Thereof as Cell Culture Media Supplement

CELL CULTURE MEDIA FOR ENHANCED PROTEIN PRODUCTION

A cell culture medium is provided which constrains cell growth and enhances antibody production. The high glucose medium of the invention is preferably saturated at 40 ~C with essential amino acids.

[BESSYVOROTOCHNAYA] CULTURAL MEDIUM FOR OBTAINING RECOMBINANT GONADOTROPINS

NOT CONTAINING SERUM MEDIUM FOR CULTIVATION OF CELLS

SUPPLEMENTS OF EUKARYOTIC CELL CULTURE MEDIA BASED ON PRODUCTS DERIVED FROM THE MILK INDUSTRY

COMPOSITION OF SAFEGUARDING Of BODY AND USES

L'invention se rapporte à l'utilisation d'une composition comprenant au moins une globine, un protomère de globine ou une hémoglobine extracellulaire d'Annélides, une solution de stabilisation et/ou une solution de conservation d'organes, ladite composition ayant une température comprise entre 0°C et 37°C, pour préserver un organe chez un donneur décédé en état de mort cérébrale ou décédé d'arrêt cardiaque. L'invention se rapporte également à un procédé de conservation d'un organe in situ dans un donneur décédé en état de mort cérébrale ou décédé d'arrêt cardiaque à l'aide des techniques CRN ou sonde de Gillot ou toute autre technique similaire.

Hücrelerin proteinsiz veya serumsuz yerleştirilmesi için ortam

Serum free cell culture medium

A serum free cell culture medium containing less than 10 ug/ml protein is disclosed. The medium comprises a basal media supplemented with levels of 2-aminoethanol substantially higher than previously disclosed and supplemented with 2-mercaptoethanol, transferrin, insulin and free essential amino acids. Unlike other serum free media, the media disclosed herein will support the growth and long term culture of a large variety of cell types including hybridomas and tumor cell lines while maintaining both growth potential and the differentiated characteristics of the particular cell line.

METHOD FOR PRODUCING ADENOHYPOPHYSIS OR PRECURSOR TISSUE THEREOF

The invention provides a method for inducing adenohypophysis or precursor tissue thereof in vitro from human pluripotent stem cells. The method includes culturing an aggregate of human pluripotent stem cells in suspension in a medium containing a bone morphogenetic protein signal transduction pathway activating substance and a substance acting on the Shh signal pathway to obtain a human cell aggregate containing a hypothalamus neuroepithelial tissue and a surface ectoderm, and further culturing the obtained human cell aggregate containing the hypothalamus neuroepithelial tissue and the surface ectoderm in suspension in a medium containing a bone morphogenetic protein signal transduction pathway activating substance and a substance acting on the Shh signal pathway to induce formation of hypophysial placode and/or Rathke's pouch in the surface ectoderm, thus obtaining a human cell aggregate containing 1) hypothalamus neuroepithelial tissue, and 2) hypophysial placode and/or Rathke's pouch ...

CULTURE MEDIA COMPRISING N-ACYL-X-GLUTAMINE DIPEPTIDES

Disclosed herein is a cell culture media containing L-glutamine from a set of N-acylated dipeptides Acyl-X-Q, and L-glutamine from a set of other glutamine-sources Qsource in a defined molar ratio R=n(Acyl-X-Q)/n(Qsource), wherein the variables X, Q, Acyl, R, n(Acyl-X-Q) and n(Qsource) are defined in the general disclosure. Processes of using the cell culture media are also described herein. 1. A cell culture media , comprisingL-glutamine from a set of N-acylated dipeptides Acyl-X-Q; andL-glutamine from a set of other glutamine-sources Qsource in a defined molar ratio R=n(Acyl-X-Q)/n(Qsource),wherein:X is defined as an L-amino acid;Q is defined as L-glutamine attached via an amide bond to L-amino acid X;Acyl is defined as a C1-C7-acyl moiety attached via an amide bond to the amino-terminus of L-amino acid X;R is defined to be in the range of 0,03 to 20;n(Acyl-X-Q) is the total amount of substance of L-glutamine contained in the set of N-acylated dipeptides Acyl-X-Q in the culture media;n(Qsource) is the total amount of substance of L-glutamine contained in the set of other glutamine sources Qsource in the culture media; andthe constituents of the set of other L-glutamine sources (Qsource) are selected from the following: Free L-glutamine, dipeptides Y-Q, or mixtures thereof, wherein Y is defined as one of the 20 genetically encoded L-amino acids, wherein Q is defined as L-glutamine attached via an amide bond to L-amino acid Y, and the amide bond connecting Y and Q in dipeptide Y-Q is a regular backbone amide bond involving the carboxy terminus of amino acid Y and the amino terminus of glutamine Q.2. The cell culture media according to claim 1 , wherein the cell culture media are serum free.3. The cell culture media according to claim 1 , wherein the cell culture media are chemically defined.4. The cell culture media according to claim 1 , wherein the only constituent of the set of other L-glutamine sources Qsource is dipeptide Y-Q claim 1 , wherein Y is Alanine.5. ...

FEED MEDIA

The invention provides stable feed media containing pyruvate and methods for stabilizing feed media by adding pyruvate. The invention further provides methods for producing proteins using such media and proteins produced through the use of such methods. 1. A method for stabilizing a concentrated feed medium for feeding a mammalian cell culture comprising including in the feed medium at least about 9 mM pyruvate ,wherein the feed medium comprises cysteine and/or cystine, wherein the sum of the concentrations of cysteine and/or cystine is at least about 7.9 mM,wherein the pH of the feed medium is from about 5.8 to about 7.4,wherein the feed medium comprises tyrosine and the tyrosine concentration is not more than about 4.4 mM, andwherein the feed medium with the included pyruvate is stable for at least about 1 week at room temperature.2. The method of claim 1 , comprising including in the feed medium at least about 18 mM pyruvate.3. The method of claim 2 , comprising including in the feed medium at least about 30 mM pyruvate.4. The method of claim 1 , wherein the pH of the feed medium is from about 6.0 to about 7.2.5. The method of claim 1 , wherein the feed medium comprises at least about 6.0 mM cysteine.6. The method of claim 5 , wherein the feed medium comprises at least about 12.0 mM cysteine.7. The method of claim 6 , wherein the feed medium comprises not more than about 40 mM cysteine.8. The method of claim 1 , wherein the feed medium comprises at least about 3 mM tyrosine.9. The method of claim 8 , wherein the feed medium comprises at least about 4 mM tyrosine.10. The method of claim 1 , wherein the feed medium is serum free.11. The method of claim 1 , wherein the feed medium comprises a protein hydrolysate.12. The method of claim 1 , wherein the feed medium has an osmolarity from about 200 mOsm to about 1000 mOsm.13. The method of claim 12 , wherein the feed medium has an osmolarity from about 500 mOsm to about 1000 mOsm.14. The method of claim 1 , wherein the ...

培地添加剤及びその利用

ПРОЦЕССЫ КУЛЬТИВИРОВАНИЯ В СИСТЕМЕ ПОСЕВНЫХ ФЕРМЕНТЕРОВ И СПОСОБЫ ИХ ПРИМЕНЕНИЯ

УСОВЕРШЕНСТВОВАНИЯ КУЛЬТУРЫ КЛЕТОК

... 1. Бессывороточная среда культуры клеток, содержащая Часть А, Часть В и Часть С, гдеa) Часть А состоит по существу из модифицированной базальной среды, в которой исключены следующие компоненты: бикарбонат натрия, буфер, одноосновный фосфат натрия, двухосновный фосфат натрия, регулятор осмолярности, поверхностно-активное вещество и моносахарид глюкозы;b) Часть В состоит по существу из неорганического источника железа; иc) Часть С содержит рекомбинантный фактор роста; буфер; регулятор осмолярности; источник энергии и по меньшей мере два различных гидролизата продуктов не животного происхождения.2. Среда культуры клеток по п. 1, где- Часть А дополнительно содержит ионы цветных металлов, витамины или их комбинацию, или- неорганическим источником железа Части В является цитрат трехвалентного железа, или- рекомбинантный фактор роста Части С выбран из группы, состоящей из инсулина или рекомбинантного аналога, IGF-1, и комбинации инсулина и IGF-1, или- буфером, который исключен из модифицированной ...

Serumfreies Medium

"VOLLSYNTHETISCHES ZELLKULTURMEDIUM"

Serumfreies Medium zur Züchtung von Tierzellen

MEDIUM FOR PROTEIN AND SERUM-FREE THE CELL CULTURE

SERUM-FREE CULTURALCENTRALADDED AND YOUR USE.

SERUM-FREE MEDIUM FOR THE PRODUCTION OF RETROVIRALEN VECTORS

FOR HCV INFECTION SUITABLE COMPOSITIONS AND PROCEDURE

METHODS AND COMPOSITIONS FOR EXPANDING AND DIFFERENTIATING INSULIN-PRODUCING CELLS

Cell culture method and utilization of the same

Methods and compositions for making antibodies and antibody derivatives with reduced core fucosylation

The invention provides methods and compositions for preparing antibodies and antibody derivatives with reduced core fucosylation.

Serum-Free Chemically Defined Cell Culture Medium

Embodiments of chemically defined cell culture media containing nutrients and growth factors free of any serum for culturing cells such as mesenchymal stem cells and methods of using embodiments of the cell culture medium for expanding cell populations such as mesenchymal stem cells while maintaining a pluripotent phenotype and methods of inducing chondrogenesis and osteogenesis of mesenchymal stem cells by admixing differentiation factors into embodiments of the cell culture medium. 1. A serum-free chemically defined cell culture medium , comprising:a) a base medium, wherein said base medium is serum-free; and i) insulin;', 'ii) one or more phospholipid growth factors; and', 'iii) one or more WNT signaling pathway activators., 'b) a base medium supplement, including2. The serum-free chemically defined cell culture medium of claim 1 , wherein said base medium comprises an amount of DMEM and an amount of MCDB having a ratio of DMEM to MCDB in the range of about 0.75:1.25 v/v to about 1.25:0.75 v/v.3. (canceled)4. The serum-free chemically defined cell culture medium of claim 2 , further including one or more cell growth factors selected from the group consisting of: bFGF claim 2 , PDGF-bb claim 2 , EGF claim 2 , TGF-beta1 and IGF-1.5. The serum-free chemically defined cell culture medium of claim 1 , wherein said insulin has a concentration in said serum-free chemically defined cell culture medium in a range of about 4 mg/L and about 6 mg/L.6. The serum-free chemically defined cell culture medium of claim 4 , wherein each of said bFGF claim 4 , PDGF-bb claim 4 , EGF claim 4 , TGF-beta1 and IGF-1 has a concentration in said serum-free chemically defined cell culture medium in a range of about 1 ng/mL to about 20 ng/mL.7. (canceled)8. The serum-free chemically defined cell culture medium of claim 2 , wherein said one or more lipid growth factors is selected from the group consisting of: LPA claim 2 , and S1P.9. The serum-free chemically defined cell culture medium of ...

Methods of culturing cells in a medium comprising transforming growth factor beta 1 and basic fibroblast growth factor

The present invention is of methods of establishing and propagating human embryonic stem cell lines using feeder cells-free, xeno-free culture systems and stem cells which are capable of being maintained in an undifferentiated, pluripotent and proliferative state in culture which is free of xeno contaminants and feeder cells.

MEDIUM FOR THE PROTEIN-FREE AND SERUM-FREE CULTIVATION OF CELLS

A medium is described for the protein-free and serum-free cultivation of cells, especially mammalian cells, whereby the medium contains a proportion of soy hydrolysate. 1. An animal protein-free and serum-free cell culture medium , the medium comprising a soy hydrolysate having a total nitrogen content of between 7.6% and 11.4% , and the medium comprising0.001-1 g/L L-asparagine,0.001-1 g/L L-cysteine,0.001-1 g/L L-cystine,0.001-1.5 g/L L-proline,0.001-1 g/L L-tryptophan, and0.05-1 g/L L-glutamine.2. The animal protein-free and serum-free cell culture medium of claim 1 , wherein the medium comprises an endotoxin content of <500 U/g.3. The animal protein-free and serum-free cell culture medium of claim 1 , wherein the medium comprises more than 10 wt. % ultrafiltered soy hydrolysate based on the total dry weight of the medium claim 1 , and wherein at least 40% of the soy hydrolysate has a molecular weight of ≦500 daltons.4. The animal protein-free and serum-free cell culture medium of claim 3 , wherein at least 50% of the soy hydrolysate has a molecular weight of ≦500 daltons.5. The animal protein-free and serum-free cell culture medium of claim 3 , wherein at least 55% of the soy hydrolysate has a molecular weight of ≦500 daltons.6. The animal protein-free and serum-free cell culture medium of claim 1 , wherein the medium contains ultrafiltered soy hydrolysate.7. The animal protein-free and serum-free cell culture medium of claim 1 , wherein the medium comprises an amino acid.8. The animal protein-free and serum-free cell culture medium of claim 7 , wherein the amino acid is selected from the group consisting of L-asparagine claim 7 , L-cysteine claim 7 , L-cystine claim 7 , L-proline claim 7 , L-tryptophan claim 7 , L-glutamine and mixtures thereof.9. The animal protein-free and serum-free cell culture medium of claim 1 , wherein the animal protein-free and serum-free cell culture medium further comprises: 1 to 100 g/L synthetic minimal medium; 0.05-1 g/L glutamine ...

METHODS FOR INCREASING MANNOSE CONTENT OF RECOMBINANT PROTEINS

The present invention relates to methods of modulating the mannose content of recombinant proteins. 1. A method for modulating mannose 5 on a recombinant protein during a mammalian cell culture process comprising limiting the amount of glucose in the cell culture medium , wherein the concentration of the glucose is from about 0 to 6 g/L , and supplementing the cell culture medium with galactose or sucrose , wherein the concentration of galactose is from 6-13 g/L or the concentration of sucrose is from about 16-24 g/L.2. The method according to claim 1 , wherein the glucose concentration in the cell culture medium is sufficient to result in a concentration of glucose in the spent medium at about 0 g/L.3. (canceled)4. A method according the claim 1 , wherein the concentration of glucose in the cell culture medium is from 4 to 6 g/L.5. The method according to claim 1 , wherein the concentration of glucose in the cell culture medium is from 1 to 3 g/L.6. The method according to claim 1 , wherein the concentration of glucose in the cell culture medium is from 2 to 3 g/L.7. The method according the claim 1 , wherein the concentration of glucose in the cell culture medium is 2.5 g/L.8. The method according the claim 1 , wherein the concentration of glucose in the cell culture medium is 0 g/L.9. (canceled)10. The method according to claim 1 , wherein the concentration of galactose is from 10 to 13 g/L.11. The method according to claim 1 , wherein the concentration of galactose in the cell culture medium is from 10 to 12 g/L.12. The method according to claim 1 , wherein the concentration of galactose in the cell culture medium is 11.5 g/L.13. (canceled)14. (canceled)15. The method according to claim 1 , wherein the limiting amount of glucose is added during a production phase.16. (canceled)17. The method according the claim 1 , wherein the cell culture process is a perfusion process.1863.-. (canceled)64. The method according to claim 1 , wherein the mammalian cells are ...

METHODS OF IDENTIFYING THERAPEUTIC TARGETS FOR TREATING ANGIOGENESIS

Provided herein is a method for assessing angiogenic effects of a test composition, the method including: providing human microvessel (MV) fragments selected to correspond to a desired patient profile; embedding the human MV fragments in a gel matrix of a three dimensional (3D) in vitro culture; providing serum free media to the 3D in vitro culture; contacting the 3D in vitro culture comprising embedded human MV fragments with a test composition; and assessing the angiogenic effects of the test composition by measuring at least one angiogenic growth parameter of the 3D in vitro culture comprising embedded human MV fragments. Also provided herein are 3D in vitro cultures useful in the disclosed methods. 1. A method for assessing angiogenic effects of a test composition , the method comprising:providing human microvessel fragments selected to correspond to a desired patient profile;embedding the human microvessel fragments in a gel matrix of a three dimensional (3D) in vitro culture;providing a serum free medium to the 3D in vitro culture comprising embedded human microvessel fragments;contacting the 3D in vitro culture comprising embedded human microvessel fragments with a test composition; andassessing the angiogenic effects of the test composition by measuring at least one angiogenic growth parameter of the 3D in vitro culture comprising embedded human microvessel fragments.2. The method of claim 1 , wherein the desired patient profile comprises a shared underlying condition or trait.3. The method of claim 1 , wherein the desired patient profile comprises a heterogeneous selection of patients.4. The method of claim 1 , wherein the gel matrix comprises collagen.5. The method of claim 1 , wherein the serum free media is selected for low angiogenic growth conditions.6. The method of claim 1 , wherein the serum free media is selected for medium angiogenic growth conditions.7. The method of claim 1 , wherein the serum free media is selected for high angiogenic growth ...

SERUM-FREE FREEZING MEDIUM USED IN ADIPOSE-DERIVED STEM CELLS AND ESTABLISHMENT OF ADIPOSE-DERIVED STEM CELL LIBRARY

Disclosed is a serum-free freezing medium used in adipose-derived stem cells and a method for establishing an adipose-derived stem cell library. The serum-free freezing medium comprises a serum-free culture medium, dimethyl sulfoxide and a serum substitution component KSR; the defects of unstable freezing quality of the adipose-derived stem cells and influence of harmful factors in serum on the adipose-derived stem cells are solved, and the adipose-derived stem cells stored have the advantages of high survival percentage, well adherence growth and strong differentiation capacity. 1. A serum-free freezing medium comprising the following ingredients: serum-free culture medium , dimethylsulfoxide (DMSO) and serum substitute Knockout™ Serum Replacement (KSR) , and the freezing medium does not contain any serum.2. The serum-free freezing medium of claim 1 , wherein claim 1 , based on the volume of serum-free freezing medium claim 1 , the content of serum-free culture medium is a and a is from 5%-15% (v/v) claim 1 , the content of DMSO is b and b is from 8%-20% (v/v) claim 1 , and the content of KSR is c and c is from 70%-85% (v/v) claim 1 , while a+b+c≦100%.3. A use of the serum-free freezing medium of for long-term storage of adipose-derived stem cells and/or establishment of an adipose-derived stem cell library.4. An adipose-derived stem cells mixture claim 1 , wherein the mixture comprises:adipose-derived stem cells, and{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'the serum-free freezing medium of .'}5. A adipose-derived stem cell library claim 4 , wherein the library comprises the adipose-derived stem cells mixture of .6. The adipose-derived stem cell library of claim 5 , wherein the mixture of adipose-derived stem cells and serum-free freezing medium is preserved in liquid nitrogen after it is cooled in a programmed gradient cooling.7. The adipose-derived stem cell library of claim 5 , wherein the recovered adipose-derived stem cells of the library have an ...

CELL CULTURE MEDIA COMPOSITION AND METHODS OF PRODUCING THEREOF

A serum free cell culture media, wherein the media is adapted to be conditioned by culturing a first set of eukaryotic cells in the media, wherein the first set of eukaryotic cells use an expression vector to excrete levels of desired complex proteins into the media; wherein said desired complex proteins include human Growth Hormone (hGH), Growth Hormone-like growth factors, insulin-like growth factors, insulin, modified insulins, cytokines, mitogenic proteases and mixtures thereof; and wherein the media is adapted to grow a set of eukaryotic cells.

Method for culturing cells

A method of culturing a cell is provided comprising the step of contacting said cell with a cell culture medium comprising at least one of FGF, VEGF, or HGF. Also provided is a cell culture medium for culturing a cell comprising at least one of FGF, VEGF, or HGF; a kit for use in the method and a cell produced by the method.

Method for cell culture

A method for stem or progenitor cell culture. More precisely, the invention relates to a method for cell culture using one or more IαI (inter-alpha trypsin inhibitor or inter-alpha inhibitor) protein(s) or part(s) thereof as a component in a cell culture media or a coating on a cell culture surface material. Furthermore the invention relates to a cell culture media and a cell culture coating/matrix provided with one or more IαI proteins(s) or part(s) thereof.

Large scale cell culture system for making meat and associated products

Provided is a large-scale cell culture system for producing products without harming animals. Also provided is a method for making meat products using this cell culture system. Further provided is a method for making personal care products using this cell culture system, as well as a method for making nutritional supplements using this cell culture system.

COMPOSITIONS AND METHODS FOR ENHANCING CELL CULTURE

Provided herein are improvements in cell culture methods and compositions related thereto. In partial particular, provided herein are compositions and methods, and kits increasing the cellular division times and viability. Also provided herein are compositions and method for performing electroporation where high levels of electroporation efficiency are achieved and where deleterious effect of electroporation on cells are decreased. 1. A method for preparing a serum free cell culture medium , the method comprising adding a lipoprotein particle composition or a lipoprotein composition to a basal culture medium , wherein the lipoprotein particle composition or a lipoprotein composition is added in an amount to function as a serum replacement.2. (canceled)3. The method of claim 1 , wherein the lipoprotein particle composition comprises lipoprotein particles obtained from human blood.410.-. (canceled)11. A serum free cell culture medium comprising one or more lipoprotein compound made by the method of claim 1 , wherein the serum free cell culture medium supports the expansion of mammalian cells and wherein the expansion of the mammalian cells is increased by at least 10% in the serum free cell culture medium comprising the one or more lipoprotein compound as compared to the same cell expanded in culture medium without the one or more lipoprotein compound but containing serum.1213.-. (canceled)14. The serum free cell culture medium of claim 11 , wherein at least one of the one or more lipoprotein compound is a component of a lipoprotein particle.1518.-. (canceled)19. The serum free cell culture medium of claim 11 , wherein the increase in cell viability is in the range of from 10% to about 75%.2021.-. (canceled)22. The serum free cell culture medium of claim 11 , wherein the mammalian cells are immune cells.2325.-. (canceled)26. A method for expanding a mammalian cell claim 1 , the method comprising incubating the mammalian cell in a serum free cell culture medium ...

Methods for increasing mannose content of recombinant proteins

The present invention relates to methods of modulating the mannose content of recombinant proteins.

THERAPEUTIC PREPARATIONS OF GAMMA-DELTA T CELLS AND NATURAL KILLER CELLS AND METHODS FOR MANUFACTURE AND USE

Provided are methods of making innate immune cell compositions containing gamma.delta (γδ) T cells and/or Natural Killer (NK) cells, and the resulting compositions and related products of manufacture and kits for use in cancer and infectious disease therapy. The methods provided herein permit tailoring of the relative amounts of gamma.delta (γδ) T cells and Natural Killer (NK) cells in the compositions, for cellular therapies against a wide variety of cancers and infectious diseases. The resulting compositions can further be used to generate compositions containing either NK cells alone or gamma.delta T cells alone, for immune cellular therapies. The compositions provided herein also can be genetically altered: the gamma delta T cells and Natural Killer cells are modified to express chimeric antigen receptors (CARS) or exogenous T cell receptors (TCRs), which can be used to target any cell surface molecule either directly or indirectly, e.g., a marker on a cancer cell or an infected cell. 1. A method for manufacturing a composition comprising a population of cells enriched in NK cells and gamma.delta T cells , comprising:obtaining a sample containing cells from a subject;depleting alpha.beta T cells from the sample under conditions that generate a depleted cell population comprising NK cells and gamma.delta T cells;exposing the depleted cell population to activation conditions comprising contacting the depleted cell population with: (a) at least one exogenous polypeptide that immunospecifically binds to a cell adhesion polypeptide, and (b) at least one exogenous polypeptide that immunospecifically binds to a different polypeptide than the cell adhesion polypeptide, wherein the different polypeptide immunospecifically binds to a NK cell activation receptor, a gamma.delta T cell activation receptor, or both a NK cell activation receptor and a gamma.delta T cell activation receptor expressed on the surface of one or more cells of the sample population; andexposing the ...

Serum-free medium

A serum-free medium for the growth of mesenchymal stem cells comprises FGF, TGF-β and lipoprotein.

NATIVE WHARTON'S JELLY STEM CELLS AND THEIR PURIFICATION

Noncultured Wharton's Jelly stem cells and methods of their purification, storage and use are provided. 130-. (canceled)31. A cell culture medium comprising a cell-depleted Wharton's Jelly filtrate , wherein the filtrate is prepared by a method comprising mincing umbilical cord tissue comprising Wharton's Jelly , subsequently diluting the umbilical cord tissue , subsequently filtering the umbilical cord tissue to generate a filtrate , and sedimenting Wharton's Jelly stem cells from the filtrate. This application is a continuation of U.S. patent application Ser. No. 13/701,329, filed Nov. 30, 2012, which is the U.S. national stage of international patent application PCT/US2011/038710, filed Jun. 1, 2011, which claims the benefit of and priority to U.S. Provisional Application No. 61/350,303, filed Jun. 1, 2010, the disclosures of which are hereby incorporated by reference in their entireties.Umbilical cord tissue is a rich source of stem cells. Blood from the umbilical cord includes stem cells, including hematopoietic stem cells that can be used to repopulate a person's blood and immune system. Wharton's Jelly, a gelatinous substance within the umbilical cord, contains an additional population of stem cells, distinct from those found in cord blood. As used herein, “Wharton's Jelly” can further include the amniotic epithelial layer of the umbilical cord. Processing and culturing the Wharton's jelly permits the isolation of mesenchymal stem cells that can be used to regenerate a variety of tissues (see, for example, U.S. Pat. No. 5,919,702).The present inventors have discovered that the process of culturing cells from Wharton's Jelly substantially changes the characteristics of the cells. Compared to a population of cells cultured in vitro, uncultured Wharton's Jelly cells are molecularly different as can be seen, for example, in a different molecular profile on their cell surfaces. More importantly, the inventors have found that minimally manipulated Wharton's Jelly ...

Modulation of Angiogenesis

The invention provides methods for treating pathological conditions that can be improved by providing angiogenesis. The invention is generally directed to provide angiogenesis by administering cells that express and/or secrete one or more pro-angiogenic factors. The invention is also directed to drug discovery methods to screen for agents that modulate the ability of the cells to express and/or secrete one or more pro-angiogenic factors. The invention is also directed to cell banks that can be used to provide cells for administration to a subject, the banks comprising cells having desired levels of expression and/or secretion of one or more pro-angiogenic factors. 1. A method for providing angiogenesis in a subject , said method comprising selecting cells that have a desired potency for expression and/or secretion of one or more pro-angiogenic factors; assaying said cells for a desired potency for expression and/or secretion of one or more pro-angiogenic factors; and administering said cells having the desired potency for expression and/or secretion of one or more pro-angiogenic factors to said subject in a therapeutically effective amount and for a time sufficient to achieve a therapeutic result , the cells being non-embryonic stem , non-germ cells that express one or more of oct4 , telomerase , rex-1 , or rox-1 and/or can differentiate into cell types of at least two of endodermal , ectodermal , and mesodermal germ layers.213-. (canceled) The invention provides methods for treating pathological conditions that can be improved by providing angiogenesis. The invention is generally directed to providing angiogenesis by administering cells that express and/or secrete one or more pro-angiogenic factors. The invention is also directed to drug discovery methods to screen for agents that modulate the ability of the cells to express and/or secrete one or more pro-angiogenic factors. The invention is also directed to cell banks that can be used to provide cells for ...

Method for detecting or separating/obtaining circulating tumor cell employing cell proliferation method

A method for detecting or separating/obtaining CTC, which is capable of reliably and stably detecting or separating/obtaining a circulating tumor cell and a circulating tumor stem cell present in trace amounts in a biological circulating fluid such as blood or lymph, even in the state where the cancer type of the tumor cells cannot be determined yet and the state where the tumor cells are present in trace amounts in the biological circulating fluid. The method is attained by detecting or separating/obtaining CTC and/or CTSC in a biological circulating fluid, comprising treatment steps (1) to (4): (1) pretreating a sample from the biological circulating fluid to obtain a mononuclear cell phase; (2) providing a well plate in which a culture medium consisting of a serum-free cell growth medium for circulating tumor cell and/or circulating tumor stem cell has been injected, and seeding thereto the mononuclear cells obtained in step (1), followed by incubation; (3) removing the culture medium from a well of the plate obtained by the incubation in step (2); and (4) detecting or separating/obtaining an adherent tumor cell attached to the well of the plate after step (3).

Cell culture improvements

The invention describes improved methods and compositions for producing a recombinant protein, e.g., an antibody, in mammalian cell culture. In addition, the invention provides improved cell culture media, including improved production media, feed solutions, and combination feeds, which may be used to improve protein productivity in mammalian cell culture.

FEED MEDIA

The invention provides stable feed media containing pyruvate and methods for stabilizing feed media by adding pyruvate. The invention further provides methods for producing proteins using such media and proteins produced through the use of such methods. 1. A method for stabilizing a concentrated feed medium for feeding a mammalian cell culture comprising including in the feed medium at least about 9 mM pyruvate ,wherein the feed medium comprises cysteine and/or cystine, wherein the sum of the concentrations of cysteine and/or cystine is at least about 7.9 mM,wherein the pH of the feed medium is from about 5.8 to about 7.4,wherein the feed medium comprises tyrosine and the tyrosine concentration is not more than about 4.4 mM, andwherein the feed medium with the included pyruvate is stable for at least about 1 week at room temperature.2. The method of claim 1 , comprising including in the feed medium at least about 18 mM pyruvate.3. The method of claim 2 , comprising including in the feed medium at least about 30 mM pyruvate.4. The method of claim 1 , wherein the pH of the feed medium is from about 6.0 to about 7.2.5. The method of claim 1 , wherein the feed medium comprises at least about 6.0 mM cysteine.6. The method of claim 5 , wherein the feed medium comprises at least about 12.0 mM cysteine.7. The method of claim 6 , wherein the feed medium comprises not more than about 40 mM cysteine.8. The method of claim 1 , wherein the feed medium comprises at least about 3 mM tyrosine.9. The method of claim 8 , wherein the feed medium comprises at least about 4 mM tyrosine.10. The method of claim 1 , wherein the feed medium is serum free.11. The method of claim 1 , wherein the feed medium comprises a protein hydrolysate.12. The method of claim 1 , wherein the feed medium has an osmolarity from about 200 mOsm to about 1000 mOsm.13. The method of claim 12 , wherein the feed medium has an osmolarity from about 500 mOsm to about 1000 mOsm.14. The method of claim 1 , wherein the ...

Cell culture medium

The invention relates to serum-free cell culture medium, wherein the medium either contains maltose as the sole carbohydrate source, or contains maltose and at least one other saccharide as carbohydrate sources. In a preferred embodiment, the additional saccharide is a monosaccharide, preferably glucose, and the medium comprises DMEM-F12. In a preferred embodiment the cells to be cultured may be CHO or HEK293 cells. Also disclosed are methods of growing and/or culturing a cell using the cell culture medium as described herein, methods of increasing protein yield using the cell culture medium as described herein, and kits thereof.

MAMMALIAN CELL CULTURE

The invention provides a method for culturing mammalian cells. The method provides greater control over cell growth to achieve high product titer cell cultures. 111-. (canceled)12. A method of culturing mammalian cells expressing a recombinant protein comprising;{'sup': 6', '6, 'establishing a mammalian cell culture in a serum-free culture medium in a bioreactor by inoculating the bioreactor with at least 0.5×10to 3.0×10cells/ml in a serum-free culture medium;'}growing the mammalian cells during a growth phase and supplementing the culture medium with bolus feeds of a serum-free feed medium,{'sup': '6', 'starting perfusion when viable cell density (VCD) is at least 10×10viable cells/mL, and'}maintaining the mammalian cells by perfusion with a serum-free perfusion medium, wherein the packed cell volume during the production phase is less than or equal to 35%.13. The method according to claim 12 , wherein perfusion begins on or about day 5 to on or about day 9 of the cell culture.14. The method according to claim 12 , wherein perfusion begins on or about day 5 to on or about day 7 of the cell culture.15. The method according to claim 12 , wherein perfusion begins when the cells have reached a production phase.16. The method according to claim 12 , wherein perfusion takes place prior to a production phase.17. The method according to claim 12 , further comprising perfusion with a serum-free perfusion medium having an L-asparagine concentration of 5 mM or less.18. The method according to claim 12 , further comprising a temperature shift.19. The method according to claim 12 , further comprising a temperature shift claim 12 , wherein the temperature is lowered from between 35° C. and 38° C. to between 30° C. and 34° C.2026-. (canceled)27. The method according to claim 12 , wherein the packed cell volume is less than or equal to 30%.2829-. (canceled)30. The method according to claim 12 , wherein perfusion comprises continuous perfusion.3138-. (canceled)39. The method ...

Methods of expanding embryonic stem cells in a suspension culture

A method of expanding and maintaining human embryonic stem cells (ESCs) in an undifferentiated state by culturing the ESCs in a suspension culture under culturing conditions devoid of substrate adherence is provided. Also provided are a method of deriving ESC lines in the suspension culture and methods of generating lineage-specific cells from ESCs which were expanded in the suspension culture of the present invention.

Cell Culture Medium for Eukaryotic Cells

Cell culture media are provided herein as are methods of using the media for cell culture and protein production from cells. 1. A cell culture medium for eukaryotic cells expressing Aflibercept , comprising:a basal medium or a feed medium; and5-methylthioadenosine or nicotinamide.2. The cell culture medium of claim 1 , wherein a concentration of 5-methylthioadenosine in the cell culture medium is at least about 10 nM.3. The cell culture medium of claim 1 , wherein a concentration of the nicotinamide in the cell culture medium is at least about 50 nM.4. The cell culture medium of further comprising one or more acids selected from lactic acid claim 1 , phenyl lactic acid claim 1 , indolelactic acid claim 1 , succinic acid claim 1 , alpha-hydroxyisovaleric acid claim 1 , alpha-hydroxyisocaproic acid claim 1 , 2-(4-hydroxyphenyl)lactic acid claim 1 , or 2-hydroxy-3-methylvaleric acid claim 1 , salts of these acids claim 1 , esters of these acids and combinations thereof.5. The cell culture medium of claim 2 , wherein a titer of the Aflibercept produced in the cell culture medium is at least about 2% greater than another cell culture medium that does not have at least about 10 nM 5-methylthioadenosine.6. The cell culture medium of claim 3 , wherein a titer of the Aflibercept produced in the cell culture medium is at least about 2% greater than another cell culture medium that does not have at least about 50 nM nicotinamide.7. The cell culture medium of claim 1 , wherein the 5-methylthioadenosine or the nicotinamide can be in the form of its salts or esters.8. The cell culture medium of claim 1 , wherein pH of the medium is maintained in the range of about 6.5 to about 8.9. The cell culture medium of claim 1 , wherein the medium does not have a protein derived from an animal.10. The cell culture medium of claim 1 , wherein the medium is a serum-free medium.11. The cell culture medium of claim 1 , wherein the medium is a chemically-defined medium.1231.-. (canceled)32. A ...

CONDITIONED MEDIUM FROM HUMAN ADULT LIVER STEM CELLS AND ITS USE IN THE TREATMENT OF LIVER DISORDERS

The invention relates to cell-free compositions obtained by culturing adult-derived human liver stem/progenitor cells (ADHLSC) in cell culture medium and isolating the resulting conditioned medium (ADHLSC-CM) that has advantageous properties, such as anti-fibrotic effects. ADHLSC-CM, compositions based on ADHLSC-CM, and other related and derived products, can be used in cell culture processes or as a medicament, more particularly for the treatment of diseases involving organ injury, organ failure, in organ or cell transplantation or the pathological disruption, inflammation, degeneration, and/or proliferation of cells within a tissue or an organ, in particular within liver. 113-. (canceled)14. A method of treating a disorder in a subject in need thereof , comprising:administering to the subject a therapeutically or prophylactically effective amount of a cell-free conditioned medium,wherein the cell-free conditioned medium is produced by culturing adult-derived human liver stem/progenitor cells (ADHLSC) in a cell culture medium and separating the cell culture medium from the cells, wherein the ADHLSC are albumin-positive, vimentin-positive, alpha smooth muscle actin-positive, cytokeratin-19-negative and CD133-negative.1516-. (canceled)17. The method of claim 14 , wherein the disorder is a fibrotic disorder.18. The method of claim 14 , wherein the disorder is a liver disorder.19. The method of claim 14 , wherein the subject is suffering from organ failure claim 14 , or is in need of organ or cell transplantation.20. A method of treating a disorder in a subject in need thereof claim 14 , comprising:administering to the subject a therapeutically or prophylactically effective amount of a fractioned cell-free composition,wherein the fractioned cell-free composition is produced by:culturing adult-derived human liver stem/progenitor cells (ADHLSC) in a cell culture medium and separating the cell culture medium from the cells to produce a cell-free conditioned medium, ...

COMPOSITIONS CONTAINING PLATELET CONTENTS

This document provides methods and materials relating to platelet lysates. For example, methods and materials for using platelet lysate compositions to grow adult stem cells, to differentiate adult stem cells, to grow primary cell cultures, to grow tumor stem cells, and to identify effective growth factors are provided. 1. A platelet lysate composition comprising a filtrate from a lysed platelet preparation passed through a 0.45 um or smaller filter and comprises greater than 200 pg of VEGF polypeptide per ml.2. The platelet lysate composition of claim 1 , wherein said filtrate is from said lysed platelet preparation being passed through a 0.45 μm filter and a 0.2 um filter.3. The platelet lysate composition of claim 1 , wherein culturing 1.4×106 mesenchymal stem cells with media containing about five percent of said platelet lysate composition results in greater than 1.4×10 7 cells after three days.4. A method for making a platelet lysate composition claim 1 , wherein said method comprises:(a) lysing platelets via one or more freeze/thaw cycles to obtain lysed platelets,(b) centrifuging said lysed platelets to obtain a supernatant, and(c) filtering said supernatant through a 0.45 μm or smaller filter to obtain a filtrate, wherein said filtrate is said platelet lysate composition.5. The method of claim 4 , wherein said lysing step comprises at least two freeze/thaw cycles.6. The method of claim 4 , wherein said centrifuging step comprises using a force between 2000×g and 4000×g for between 15 and 45 minutes.7. The method of claim 4 , wherein said filtering step comprising filtering said supernatant through a 0.45 μm filter and a 0.2 μm filter.8. A method for expanding a cell population comprising culturing a first population of cells in the presence of medium comprising a platelet lysate composition under conditions wherein said first population of cells is expanded to a second population of cells having more cells than said first population claim 4 , wherein said ...

Method for culturing mdck cells

The present invention relates to a cloned MDCK cell showing an expansion factor of 4.5 or more when cultured using a microcarrier and a method of culturing the MDCK cell, a method of growing a virus using the method of culturing the MDCK cell, and a cloned MDCK cell showing an expansion factor of 4.5 or more when cultured using a microcarrier.

METHODS OR GENERATING T-CELLS FROM STEM CELLS AND IMMUNOTHERAPEUTIC METHODS USING THE T-CELLS

Methods and composition for production of T cells are provided. Also provided are therapeutic methods using engineered T cells. For example, in certain aspects methods include preparing three dimensional cell culture compositions comprising stroma cells and hematopoietic stem or progenitor cells in a serum-free medium for producing T cells. 1186-. (canceled)187. A method for preparing a composition of mature CD4 CD8 , or CD8ab CD4 T cells from stem or progenitor cells , the method comprising culturing a three-dimensional (3D) cell aggregate comprising:a) a cell line of stromal cells that express a Notch ligand; andb) a population of stem or progenitor cells generated in vitro from embryonic stem cells (ESCs), induced pluripotent stem cells (iPSC), or human embryonic mesodermal progenitor cells;wherein the 3D cell aggregate is cultured in a serum-free medium for a time period sufficient for in vitro differentiation of the stem or progenitor cells to mature T cells, wherein the 3D cell aggregate does not comprise an exogenous matrix or a scaffold.188. The method of claim 187 , wherein the Notch ligand is an exogenous Notch ligand.189. The method of claim 187 , wherein the medium further comprises one or more of externally added FLT3 ligand (FLT3L) claim 187 , interleukin 7 (IL-7) claim 187 , stem cell factor (SCF) claim 187 , thrombopoietin (TPO) claim 187 , thrombopoietin (TPO) claim 187 , IL-2 claim 187 , IL-4 claim 187 , IL-6 claim 187 , IL-15 claim 187 , IL-21 claim 187 , TNF-alpha claim 187 , TGF-beta claim 187 , interferon-gamma claim 187 , interferon-lambda claim 187 , TSLP claim 187 , thymopentin claim 187 , pleiotrophin claim 187 , midkine claim 187 , or combinations thereof.190. The method of claim 187 , wherein the stromal cells have an exogenous nucleotide sequence encoding an intact claim 187 , partial or modified Notch ligand claim 187 , and wherein the Notch ligand is DLL4 claim 187 , DLL1 claim 187 , JAG1 claim 187 , JAG2 claim 187 , or a combination ...

Cell Culture Improvements

The invention describes improved methods and compositions for producing a recombinant protein, e.g., an antibody, in mammalian cell culture. In addition, the invention provides improved cell culture media, including improved production media, feed solutions, and combination feeds, which may be used to improve protein productivity in mammalian cell culture.

Method of expansion of human pancreas progenitor cells from stem cells using feeder-conditioned media

The present disclosure provides a method of producing and expanding human pancreas progenitor cells using, for example, iPSC derived cells and a human feeder cell conditioned medium. In one embodiment, cardiac mesenchyme cells are employed as feeder cells and those cells secrete growth factors, such as one or more of FGF10, KGF, or EGF, that promote pancreatic bud formation and expansion during development. In one embodiment, feeder cells are isolated from human stem cells, e.g., a human iPS-derived cardiac cells, and used to condition media and promote the growth and proliferation of iPSc derived pancreatic progenitor cells (in a feeder-free system).

Cell Culture Improvements

The invention describes improved methods and compositions for producing a recombinant protein, e.g., an antibody, in mammalian cell culture. In addition, the invention provides improved cell culture media, including improved production media, feed solutions, and combination feeds, which may be used to improve protein productivity in mammalian cell culture. 121-. (canceled)22. A fed batch method for making an anti-TNFα antibody comprising a light chain variable region (LCVR) comprising the sequence of SEQ ID NO:1 and a heavy chain variable region (HCVR) comprising the sequence of SEQ ID NO:2 , said method comprising culturing mammalian cells comprising a nucleic acid encoding said anti-TNFα antibody in a cell culture production medium in large scale , wherein the pH of the cell culture production medium is adjusted according to a pH linear ramp comprising beginning at a starting pH and ending at a final pH that is less than the starting pH , such that said anti-TNFα antibody is produced at a titer of at least 2 g/L in said cell culture production medium.23. The fed batch method according to claim 22 , wherein the starting pH is 8 or less.24. The fed batch method according to claim 22 , wherein the final pH is 6.5 to 7.0.25. The fed batch method according to claim 24 , wherein the starting pH is 8 or less.26. The fed batch method according to claim 22 , wherein the starting pH is 7.1 to 7.2.27. The fed batch method according to claim 22 , wherein the final pH is 6.9.28. The fed batch method according to claim 22 , wherein the starting pH is 6.5 to 8 and the final pH is 6.5 to 7.29. The fed batch method according to claim 22 , wherein the starting pH is 7.1 and the final pH is 6.9.30. The fed batch method according to claim 22 , wherein the pH is adjusted over a period of at least 24 hours claim 22 , at least 48 hours claim 22 , or at least 72 hours.31. The fed batch method according to claim 22 , wherein the pH is adjusted within the first 72 hours of the culturing.32 ...

METHODS FOR INCREASING MANNOSE CONTENT OF RECOMBINANT PROTEINS

The present invention relates to methods of modulating the mannose content of recombinant proteins.

METHODS FOR INCREASING MANNOSE CONTENT OF RECOMBINANT PROTEINS

The present invention relates to methods of modulating the mannose content of recombinant proteins. 1. A method for modulating mannose 5 on a recombinant protein during a mammalian cell culture process comprising:establishing a cell culture of mammalian cells in a cell culture medium, wherein the mammalian cells produce a recombinant protein;limiting the amount of glucose in the cell culture medium, wherein the concentration of the glucose is from about 0 to 6 g/L;and supplementing the cell culture medium with galactose or sucrose, wherein the concentration of galactose is from 6-13 g/L or the concentration of sucrose is from about 16-24 g/L in the culture medium.2. The method according to claim 1 , wherein the glucose concentration in the cell culture medium is sufficient to result in a concentration of glucose in the spent medium at about 0 g/L.3. (canceled)4. A method according the claim 1 , wherein the concentration of glucose in the cell culture medium is from 4 to 6 g/L.5. The method according to claim 1 , wherein the concentration of glucose in the cell culture medium is from 1 to 3 g/L.6. The method according to claim 1 , wherein the concentration of glucose in the cell culture medium is from 2 to 3 g/L.7. The method according the claim 1 , wherein the concentration of glucose in the cell culture medium is 2.5 g/L.8. The method according the claim 1 , wherein the concentration of glucose in the cell culture medium is 0 g/L.9. (canceled)10. The method according to claim 1 , wherein the concentration of galactose is from 10 to 13 g/L.11. The method according to claim 1 , wherein the concentration of galactose in the cell culture medium is from 10 to 12 g/L.12. The method according to claim 1 , wherein the concentration of galactose in the cell culture medium is 11.5 g/L.13. (canceled)14. (canceled)15. The method according to claim 1 , wherein the limiting amount of glucose is added during a production phase.16. (canceled)17. The method according the claim 1 , ...

CELL CULTURE MEDIUM, CULTURE METHOD, AND ORGANOID

The present invention provides a cell culture medium with which serum-free long-term culture of an epithelial stem cell, an epithelial cancer cell, or a tissue containing at least one thereof can be achieved. The cell culture medium of the present invention includes: a Wnt agonist composed of a complex of Wnt protein and afamin, which is a substance capable of stabilizing the Wnt protein, and R-spondin; and at least one selected from the group consisting of a mitogenic growth factor, a bone morphogenetic protein (BMP) inhibitor, a transforming growth factor-β (TGF-β) inhibitor, and a p38 inhibitor. 1. A cell culture medium comprising:a Wnt agonist including a complex of a Wnt protein and an afamin, which is a substance capable of stabilizing the Wnt protein; andat least one selected from the group consisting of an R-spondin, a mitogenic growth factor, a bone morphogenetic protein (BMP) inhibitor, a transforming growth factor-β (TGF-β) inhibitor, and a p38 inhibitor.2. The cell culture medium according to claim 1 , wherein the Wnt protein is at least one selected from the group consisting of Wnt1 claim 1 , Wnt2 claim 1 , Wnt2b claim 1 , Wnt3 claim 1 , Wnt3a claim 1 , Wnt4 claim 1 , Wnt5a claim 1 , Wnt5b claim 1 , Wnt6 claim 1 , Wnt7a claim 1 , Wnt7b claim 1 , Wnt8a claim 1 , Wnt8b claim 1 , Wnt9a claim 1 , Wnt9b claim 1 , Wnt10a claim 1 , Wnt10b claim 1 , Wnt11 claim 1 , and Wnt16.3. The cell culture medium according to claim 1 , wherein the R-spondin is at least one selected from the group consisting of R-spondin 1 claim 1 , R-spondin 2 claim 1 , R-spondin 3 claim 1 , and R-spondin 4.4. The cell culture medium according to claim 1 , wherein the mitogenic growth factor is an epidermal growth factor (EGF).5. The cell culture medium according to claim 1 , wherein the BMP inhibitor is Noggin.6. The cell culture medium according to claim 1 , wherein the TGF-β inhibitor is A83-01.7. The cell culture medium according to claim 1 , which is a serum-free medium.8. A method ...

CELL CULTURE MEDIUM AND CULTURE MEDIUM SUPPLEMENT

In one aspect, the present disclosure relates to a substantially albumin-free, chemically defined medium for efficiently supporting stem cell differentiation with significantly improved reproducibility and long-term culture of the differentiated cells. In particular embodiments, provided herein are compositions and methods for promoting atrial and ventricular cardiomyocytes formation from stem cells. The present disclosure further relates to the atrial and ventricular cardiomyocytes formed from the stem cells, and the uses of the cardiomyocytes, e.g., for cardiac injury repairmen, cardiac safety evaluation during drug discovery, and screening for new therapeutics for treating cardiac injuries. 1. A cell culture medium supplement comprising at least one antioxidant or at least two different antioxidants that substitute(s) the function of albumin in a cell culture medium , wherein said cell culture medium supplement is configured to be combined with a basal culture medium to form a substantially albumin-free cell culture medium.2. A cell culture medium supplement comprising at least one antioxidant or at least two different antioxidants selected from the group consisting of:a) ascorbic acid, ascorbate, or a salt or an ester thereof,b) a water-soluble analog of vitamin E,c) N-acetyl-cysteine or glutathione, or a salt or an ester thereof,d) pyruvic acid, pyruvate, or a salt or an ester thereof,e) a catalase,f) a superoxide dismutase,g) a thiol, such as 2-mercaptoethanol or 1-thioglycerol,h) a metallothione,i) a thioredoxin,j) lipoic acid or a salt or an ester thereof,k) uric acid or a salt or an ester thereof,l) a carotene,m) melatonin,n) probucol,o) dimethylthiourea, andp) resveratrol,wherein said cell culture medium supplement is configured to be combined with a basal culture medium to form a substantially albumin-free cell culture medium.38-. (canceled)9. The cell culture medium supplement of claim 1 , which further comprises an iron carrier.10. The cell culture ...

CORN ACTIVE PEPTIDE ADDITIVE FOR CELL CULTURE MEDIUM

The present invention provides a corn active peptide additive for cell culture medium, wherein in the corn active peptide additive, oligopeptides with molecular weight of lower than 1000 Dalton account for equal to or more than 90 wt % of total proteins, and the oligopeptides at least comprise one or more of AP, SAP, PAL, VNAP, PSSQ, and TQPGPQ. The corn active peptide additive of the present invention can be compounded with various basic culture mediums for serum-free culture of various animal cells, which not only substantially lowers the cost for cell culturing and reduces pollution and other problems caused by an animal derived component, but also can promote cell proliferation, improve cell viability and enhance expression of cell products. 1. A corn active peptide additive for cell culture medium , wherein in the corn active peptide additive , oligopeptides with molecular weight of lower than 1000 Dalton account for equal to or more than 90 wt % of total proteins , and the oligopeptides at least comprise one or more of AP , SAP , PAL , VNAP , PSSQ , and TQPGPQ.2. The corn active peptide additive according to claim 1 , wherein the oligopeptide further comprises one or more of AY claim 1 , NAP claim 1 , PVIN claim 1 , and AYPQ.3. The corn active peptide additive according to claim 1 , wherein endotoxin content in the corn active peptide additive is less than 200 EU/g.4. The corn active peptide additive according to claim 1 , wherein the corn active peptide additive is obtained by performing enzymolysis on corn gluten meal using a non-specific protease and a specific protease in sequence and then performing separation and purification.5. The corn active peptide additive according to claim 4 , wherein the non-specific protease is selected from one or more of papain claim 4 , alkaline protease claim 4 , neutral protease and bromelain claim 4 , and the specific protease is selected from one or two of carboxypeptidase and flavourzyme.6. A serum-free culture medium ...

CELL CULTURE MEDIUM FOR EUKARYOTIC CELLS

Cell culture media are provided herein as are methods of using the media for cell culture and protein production from cells. 1. A method for culturing eukaryotic cells for increasing production of aflibercept , comprising the steps of:culturing eukaryotic cells expressing aflibercept in culture medium;supplementing the cell culture medium with 5-methylthioadenosine, wherein a concentration of the 5-methylthioadenosine in said cell culture medium is from about 10 nM to about 200 nM; andwherein the supplementation with 5-methylthioadenosine increases a titer of said aflibercept.2. The method of claim 1 , wherein the titer of said aflibercept is at least about 2% greater than another method with a cell culture medium that has less than 10 nM of 5-methylthioadenosine.3. The method of claim 1 , wherein the cell culture medium further comprises one or more acids selected from lactic acid claim 1 , phenyllactic acid claim 1 , indolelactic acid claim 1 , succinic acid claim 1 , alpha-hydroxyisovaleric acid claim 1 , alpha-hydroxyisocaproic acid claim 1 , 2-(4-hydroxyphenyl)lactic acid claim 1 , or 2-hydroxy-3-methylvaleric acid claim 1 , salts of these acids claim 1 , esters of these acids and combinations thereof.4. The method of claim 1 , wherein the eukaryotic cells include at least one selected from the group consisting of: baby hamster kidney cell lines claim 1 , Chinese hamster ovary cell lines claim 1 , murine myeloma cell lines claim 1 , mouse myeloma cell lines claim 1 , human embryonic kidney cell lines claim 1 , human retina-derived cell lines claim 1 , and amniocyte cell lines.5. The method of claim 1 , wherein the aflibercept is secreted in the medium.6. The method of claim 1 , wherein the cell culture medium does not have a protein derived from an animal having a pH between 6.5 and 8.0.7. The method of claim 1 , wherein the cell culture medium is a serum-free medium having a pH between 6.5 and 8.0.8. The method of claim 1 , wherein the cell culture medium is a ...

SYNTHETIC ATTACHMENT MEDIUM FOR CELL CULTURE

An aqueous cell culture medium composition includes an aqueous cell culture solution configured to support the culture of mammalian cells. The composition further includes a synthetic polymer conjugated to a polypeptide dissolved in the aqueous cell culture solution. The synthetic polymer conjugated to a polypeptide is configured to attach to the surface of a cell culture article under cell culture conditions. Incubation of the aqueous cell culture medium composition on a cell culture surface under cell culture conditions results is attachment to the surface of the synthetic polymer conjugated to the polypeptide. 135-. (canceled)37. The aqueous cell culture medium composition of claim 36 , wherein the cell culture medium composition is a chemically defined composition.38. The aqueous cell culture medium composition of wherein synthetic copolymer is introduced to cell culture medium in a dry state.39. The aqueous cell culture medium composition wherein the synthetic copolymer is added to cell culture media dissolved in water.40. The aqueous cell culture medium composition of claim 36 , wherein the molar ratio of the methacrylic acid conjugated to the polypeptide and the hydroxyethylmethacrylate is between 1 to 50 and 1 to 1.41. The aqueous cell culture medium composition of wherein the synthetic copolymer is formed from copolymerization of the hydroxyethylmethacrylate and MAA-PEO4-polypeptide claim 36 , wherein MAA is the methacrylic acid claim 36 , and PEO is polyethylene oxide.42. The aqueous cell culture medium composition of claim 36 , wherein the weight percentage of the polypeptide relative to the copolymer conjugated to the polypeptide is greater than 10%.43. The aqueous cell culture medium composition of claim 36 , wherein the polypeptide is a cell adhesive polypeptide.44. The aqueous cell culture medium composition of claim 36 , wherein the polypeptide comprises an RGD sequence.45. The aqueous cell culture medium composition of claim 36 , wherein the ...

CULTURE, DEVICE, AND COMPOSITION FOR PRODUCING ADULT OLIGODENDROCYTE-TYPE 2 ASTROCYTE PROGENITOR CELLS AND PROLIFERATING OLIGODENDROCYTE-TYPE 2 ASTROCYTE PROGENITOR CELLS

The present invention provides a method for producing adult oligodendrocyte progenitor cells from proliferative oligodendrocyte progenitor cells, and a pharmaceutical composition having for an active ingredient thereof adult OPC produced according to that method. The method for producing adult OPC of the present invention is characterized by inducing proliferating OPC to differentiate into adult OPC by culturing in the presence of a ligand of a thyroid hormone receptor or retinoic acid receptor in a low oxygen environment. The present invention further provides adult OPC produced according to the production method of the present invention, and a pharmaceutical composition having these adult OPC as an active ingredient thereof. 1. A culture comprising:a serum-free medium not containing a thyroid hormone; andproliferating OPC.2. The culture according to claim 1 , wherein the proliferating OPC are a primary culture or a sub-culture of OPC collected from an organism.3. A device for producing adult OPC claim 1 , comprising:a composition comprising proliferating OPC and a serum-free medium; andan oxygen concentration controller.4. A device for producing proliferating OPC claim 1 , comprising:a composition comprising proliferating OPC and a serum-free medium not containing a thyroid hormone; andan oxygen concentration controller.5. The device for producing adult OPC according to claim 3 , the composition further comprising a ligand of a thyroid hormone receptor or retinoic acid receptor selected from the group consisting of 3 claim 3 ,5 claim 3 ,3′ claim 3 ,5′-tetraiodo-L-thyronine (T4) claim 3 , 3 claim 3 ,5 claim 3 ,3′-triiodo-L-thyronine (T3) claim 3 , a tetrazole compound having a structure similar to T4 or T3 and being able to bind to the thyroid hormone receptor claim 3 , retinoic acid claim 3 , and vitamin A.6. The device for producing proliferating OPC according to claim 4 , the composition further comprising a ligand of a thyroid hormone receptor or retinoic acid ...

METHODS FOR ENHANCED PRODUCTION AND ISOLATION OF CELL-DERIVED VESICLES AND TREATMENT OF INFLAMMATION AND NEUROLOGICAL DAMAGE

This disclosure relates to populations and compositions of purified cell-derived vesicles and uses thereof. One aspect of the disclosure relates to methods for purifying the cell-derived vesicles. 1. A method for treating a disease or condition involving an inflammatory response or related to inflammation in a subject in need thereof , comprising administering to the subject a purified population of cell-derived vesicles , wherein the population is purified from a population of stem cells cultured under conditions of hypoxia and low serum , and optionally wherein the cell-derived vesicles comprise exosomes and/or microvesicles.2. The method of claim 1 , wherein the inflammatory disease or condition is selected from multiple sclerosis claim 1 , primary and secondary progressive multiple sclerosis claim 1 , relapsing remitting multiple sclerosis claim 1 , radiation-induced soft tissue damage claim 1 , fralility claim 1 , a neuroinflammatory disease claim 1 , muscle injuries claim 1 , radiation tissue damage claim 1 , stroke claim 1 , brain inflammatory disease claim 1 , traumatic brain injury claim 1 , myocardial infarction claim 1 , graft versus host disease claim 1 , Parkinson's disease claim 1 , Alzheimer's claim 1 , inflammatory bowel disease claim 1 , Huntington's disease claim 1 , amyotrophic lateral sclerosis claim 1 , Bahcet's disease claim 1 , sarcopenia claim 1 , aging claim 1 , spinal cord injury claim 1 , wound repair claim 1 , or dysphagia claim 1 , and optionally wherein the disease or condition excludes stroke.3. The method of claim 1 , wherein the inflammatory disease or condition is selected from multiple sclerosis claim 1 , primary and secondary progressive multiple sclerosis claim 1 , or relapsing remitting multiple sclerosis.4. The method of claim 1 , wherein the subject is administered at least one dose of cell-derived vesicles selected from the group of: from between approximately 0.1 mg to about 1000 mg of cell-derived vesicle protein from the ...

COMPOSITIONS CONTAINING PLATELET CONTENTS

This document provides methods and materials relating to platelet lysates. For example, methods and materials for using platelet lysate compositions to grow adult stem cells, to differentiate adult stem cells, to grow primary cell cultures, to grow tumor stem cells, and to identify effective growth factors are provided. 118-. (canceled)19. A method for expanding a cell population for the treatment of a disease state comprising:culturing a first population of cells in the presence of medium comprising a platelet lysate composition, wherein the platelet lysate composition comprises, plasma and a total protein concentration of at least about 35 mg/mL, 200 pg or more of VEGF polypeptide per ml and wherein said platelet lysate composition comprises protein complexes greater than 50 kD.20. The method of claim 19 , wherein the cells are primary tumor cells claim 19 , tumor stem cells claim 19 , mesenchymal stromal cells claim 19 , adult stem cells claim 19 , endothelial precursor cells claim 19 , fibroblasts claim 19 , epithelial cells claim 19 , human primary glioma cells and combination thereof.21. The method of claim 19 , wherein the platelet lysate composition comprises a protein fraction having a molecular weight of greater than 100 claim 19 ,000 daltons and that contains growth factor activity.22. The method of claim 19 , wherein the platelet lysate composition comprises an average of about 180 pg/mL FGF.23. The method of claim 19 , wherein the platelet lysate composition comprises an average of about 8.5 ng/mL PDGF.24. The method of claim 19 , wherein the platelet lysate composition comprisesabout 75 to about 315 pg FGF per mL;about 4 to about 14 ng PDGF per mL;about 90 to about 155 ng IGF-1 per mL; orabout 90 to about 150 ng TGF-β per mL.25. The method of claim 19 , wherein said platelet lysate composition comprises protein complexes greater than 75 kDa.26. The method of claim 19 , wherein said platelet lysate composition comprises protein complexes greater than ...

Induction of Corneal Endothelial Cells

Compositions and methods for producing major ocular cell types, including retinal ganglion cells, photoreceptors, retinal pigmented epithelium and corneal endothelial cells, from human pluripotent stem cells under defined culture conditions are provided. 1. A method of culturing corneal endothelial cells (CECs) in vitro comprising , providing a culture of ocular neural crest stem cells (NCSCs) , and suppressing TGFβ and ROCK signaling in the culture , thereby culturing CECs.2. The method of claim 1 , wherein the suppressing step is conducted in a feeder-free and serum-free defined media by applying a small molecule differentiator to the culture.3. The method of claim 2 , wherein the small molecule differentiator is selected from a ROCK inhibitor claim 2 , a TGFβ inhibitor claim 2 , or both.4. The method of claim 3 , wherein the ROCK inhibitor is H-1125 or Y21632 claim 3 , and wherein the TGFβ inhibitor is SB431542.5. The method of claim 1 , wherein the CEC culture is expanded into a sheet of cells.6. The method of claim 1 , further comprising the earlier step of providing a culture of eye field stem cells (EFSCs) and promoting WNT signaling in the EFSCs to produce the culture of NCSCs.7. The method of claim 6 , wherein WNT signaling is promoted by removing a WNT pathway signaling inhibitor from the EFSCs.8. The method of claim 6 , wherein the majority of NCSCs express neural crest markers p75NTR and HNK-1.9. The method of claim 6 , further comprising the earlier step of providing a culture of pluripotent stem cells (PSCs) and inducing differentiation to become EFSCs.10. The method of claim 9 , wherein the majority of EFSCs express markers PAX6 and LHX2.11. The method of claim 9 , wherein PSCs are cultured with a WNT pathway signaling inhibitor claim 9 , a BMP pathway signaling inhibitor claim 9 , and a TGF-β pathway signaling inhibitor.12. The method of claim 1 , wherein the WNT pathway signaling inhibitor is IWP2 claim 1 , the BMP pathway signaling inhibitor is ...

CONDITIONED MEDIUM FROM HUMAN ADULT LIVER STEM CELLS AND ITS USE IN THE TREATMENT OF LIVER DISORDERS

The invention relates to cell-free compositions obtained by culturing adult-derived human liver stem/progenitor cells (ADHLSC) in cell culture medium and isolating the resulting conditioned medium (ADHLSC-CM) that has advantageous property ties, such as anti-fibrotic effects. ADHLSC-CM, compositions based on ADHLSC-CM, and other related and derived products, can be used in cell culture processes or as a medicament, more particularly for the treatment of diseases involving organ injury, organ failure, in organ or cell transplantation or the pathological disruption, inflammation, degeneration, and/or proliferation of cells within a tissue or an organ, in particular within liver. 1. A cell-free conditioned medium that contains soluble proteins and/or micro vesicles and that is obtainable by culturing adult-derived human liver stem/progenitor cells (ADHLSC) in a cell culture medium and separating the cell culture medium from the cells.2. The cell-free conditioned medium according to claim 1 , wherein the medium is serum-free.3. A cell-free composition obtainable by fractioning the cell-free conditioned medium according to claim 1 , wherein said fractioning comprises filtering claim 1 , enzymatically digesting claim 1 , centrifuging claim 1 , adsorbing claim 1 , and/or separating by chromatography the cell-free conditioned medium.4. The cell-free conditioned medium according to claim 1 , wherein adult-derived human liver stem/progenitor cells (ADHLSC) co-express at least one mesenchymal marker selected from CD90 claim 1 , CD73 claim 1 , CD44 claim 1 , vimentin and a-smooth muscle actin with at least an hepatic marker selected from albumin claim 1 , CD29 claim 1 , alpha-fetoprotein claim 1 , alpha-1 antitrypsin claim 1 , HNF-4 and MRP2 transporter.5. The cell-free conditioned medium according to claim 1 , wherein adult-derived human liver stem/progenitor cells (ADHLSC) are albumin-positive claim 1 , vimentin-positive claim 1 , alpha smooth muscle actin-positive claim 1 , ...

CELL FREEZING MEDIUM FOR CLINICAL USE

Provided in the present invention is a cell freezing medium for clinical use. In particular, the cell freezing medium of the present invention comprises the following components: (1) human albumin; (2) cryoprotectant: the cryoprotectant comprises a combination of one or more of dimethyl sulfoxide, glycerol, and ethylene glycol; (3) a saline buffer; wherein the salt buffer is a solution containing Na, K, Mg, Cl, and CHCOO ions; (4) a vitamin; and (5) an amino acid, wherein the human albumin concentration is 1%-20% (w/v). The cell, after long-term cryopreservation with the freezing medium of the present invention, has a high viability, and the cellular efficiency maintains a high uniformity. The grade of purity of the freezing medium of the present invention is the pharmaceutical grade or USP grade; and the freezing medium is safe and reliable for clinical use, and can be used or conventional adherent and suspension cells. 1. A cell cryopreservation solution for long-term storage of a mammalian stem cell , comprising:(a) an albumin; and(b) amino acids; wherein the amino acids consist of a combination of glycine and arginine or a combination of leucine and phenylalanine.2. The cell cryopreservation solution of claim 1 , further comprising a cryoprotectant comprising one or more of dimethyl sulfoxide claim 1 , glycerin claim 1 , and ethylene glycol.3. The cell cryopreservation solution of claim 1 , further comprising a salt buffer comprising one or more of Na claim 1 , K claim 1 , Mg claim 1 , Cland CH3COOions.4. The cell cryopreservation solution of claim 1 , further comprising a vitamin.5. The cell cryopreservation solution of claim 1 , wherein the albumin is a human albumin.6. The cell cryopreservation solution of claim 1 , wherein the albumin is present at a concentration of 1%-20% weight by volume (w/v).7. The cell cryopreservation solution of claim 5 , wherein the human albumin is selected from the group consisting of a plasma extracted human albumin claim 5 , a ...

Cell Culture Improvements

The invention describes improved methods and compositions for producing a recombinant protein, e.g., an antibody, in mammalian cell culture. In addition, the invention provides improved cell culture media, including improved production media, feed solutions, and combination feeds, which may be used to improve protein productivity in mammalian cell culture.

EX VIVO MEAT PRODUCTION

Systems and methods for producing cell cultured food products. The cultured food products include sushi-grade fish meat, fish surimi, foie gras, and other food types. Various cell types are utilized to produce the food products and can include muscle, fat, and/or liver cells. The cultured food products are grown in pathogen-free culture conditions without exposure to toxins and other undesirable chemicals. 1. A method of producing cultured tissue for human consumption , the method comprising:a) obtaining a population of self-renewing cells;b) culturing the population of self-renewing cells;c) inducing differentiation in the population of self-renewing cells to form cultured tissue; andd) processing the cultured tissue for human consumption.2. The method of claim 1 , wherein obtaining the population of self-renewing cells comprises transitioning a population of cells from 2-dimensional adherent culture into 3-dimensional culture in a bioreactor.3. The method of claim 1 , wherein the population of self-renewing cells comprises differentiated cells that have become immortalized.4. The method of any one of - claim 1 , wherein inducing differentiation in the population of self-renewing cells comprises inducing transdifferentiation of cells in the population into myocytes claim 1 , adipocytes claim 1 , or a combination thereof.5. The method of claim 1 , wherein culturing comprises seeding the population of self-renewing cells on 3-dimensional micro-scaffolds.6. The method of claim 3 , wherein the 3-dimensional micro-scaffolds promote cell growth claim 3 , adhesion claim 3 , differentiation claim 3 , or a combination thereof.7. The method of claim 3 , wherein the 3-dimensional micro-scaffolds are conjugated to at least one factor promoting cell growth claim 3 , adhesion claim 3 , differentiation claim 3 , or a combination thereof.8. The method of claim 7 , wherein the micro-scaffolds comprise at least one of hydrogel claim 7 , chitosan claim 7 , polyethylene terephthalate ...

CELL CULTURE IMPROVEMENTS

The invention describes improved methods and compositions for producing a recombinant protein, e.g., an antibody, in mammalian cell culture. In addition, the invention provides improved cell culture media, including improved production media, feed solutions, and combination feeds, which may be used to improve protein productivity in mammalian cell culture. 120-. (canceled)21. A fed-batch production method of making a human anti-TNFα antibody which comprises (1) a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:1 and (2) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:2 , said method comprising culturing Chinese Hamster Ovary (CHO) cells comprising a nucleic acid encoding said anti-TNFα antibody in a cell culture production medium , wherein the glucose concentration in said medium is monitored and glucose is added to said medium if the glucose concentration in said medium decreases to below 2 g/L , or the glucose concentration in said medium is monitored and glucose is added to said medium to maintain the glucose concentration in said medium at a concentration of at least 2 g/L , such that said anti-TNFα antibody is produced at a titer of at least 2 g/L in said cell culture production medium.22. The method of claim 21 , wherein said anti-TNFα antibody is produced at a titer of at least 2.5 g/L in said cell culture production medium.23. The method of claim 21 , wherein said anti-TNFα antibody is produced at a titer of at least 4 g/L in said cell culture production medium.24. The method of claim 21 , wherein said culturing of said CHO cells is performed in a large scale culture volume of greater than 10 L.25. The method of claim 22 , wherein said culture is performed in a large scale culture volume of greater than 10 L.26. The method of claim 23 , wherein said culture is performed in a large scale culture volume of greater than 10 L.27. The method of claim 24 , wherein said culturing lasts 9-15 days.28. ...

Compositions for treatment of osteochondral disorders

The application provides biocompatible carriers comprising bone forming and/or cartilage forming cells and methods for making them. The application further provides pharmaceutical compositions comprising said ATMPs and method of treatments using said ATMPs. The application further relates to said ATMPS for use in the treatment of bone disorders, cartilage disorders and joint disorders. The current invention further relates to method of treatments of bone disorders, cartilage disorders and joint disorders.

METHODS AND COMPOSITIONS FOR MAKING ANTIBODIES AND ANTIBODY DERIVATIVES WITH REDUCED CORE FUCOSYLATION

The invention provides methods and compositions for preparing antibodies and antibody derivatives with reduced core fucosylation. 154-. (canceled)56. The fucose analog of claim 55 , wherein each of R-Ris independently selected from the group consisting of —OH and —OC(O)C-Calkyl.57. The fucose analog of claim 55 , wherein each of R-Ris independently selected from the group consisting of —OH and —OAc.58. The fucose analog of claim 55 , wherein Ris selected from the group consisting of —C≡CH and —C≡CCH.59. The fucose analog of claim 55 , wherein Ris —C(O)OCH.60. The fucose analog of claim 55 , wherein Ris selected from the group consisting of —CN and —CHCN.61. The fucose analog of claim 55 , wherein Ris —CHCN.62. The fucose analog of claim 55 , wherein Ris —CHBr claim 55 , —CHCl claim 55 , or —CHI.63. The fucose analog of claim 55 , wherein Ris —CHBr or —CHI.64. The fucose analog of claim 55 , wherein Ris —CHBr.65. The fucose analog of claim 55 , wherein Ris methoxiran.66. The fucose analog of claim 55 , wherein Ris —CH(OAc)CH.6772-. (canceled)74. A compound of claim 73 , wherein Ris F.75. A compound of claim 73 , wherein Ris F.76. A compound of claim 73 , wherein Ris F.77. A compound of claim 73 , wherein Ris F and Ris F.78. A compound of claim 73 , wherein each of Rand Ris F.79. A compound of claim 73 , wherein R claim 73 , Rand Rare each independently selected from the group consisting of —OH and —OAc; Ris F; and Ris —CH.80. A compound of claim 73 , wherein R claim 73 , Rand Rare each independently selected from the group consisting of —OH and —OAc; Ris F; Rand Rare each H; and Ris —CH.8194-. (canceled)95. (canceled) This application is a continuation of U.S. patent application Ser. No. 14/632,925 filed Feb. 26, 2015, which is a continuation of U.S. patent application Ser. No. 14/043,742 filed Oct. 1, 2013 (now U.S. Pat. No. 8,993,326), which is a divisional of Ser. No. 13/405,143 filed Feb. 24, 2012 (now U.S. Pat. No. 8,574,907), which is a divisional of Ser. No. ...

CULTURE MEDIUM FOR USE IN DIFFERENTIATION OF PLURIPOTENT STEM CELL INTO NEURAL STEM CELL, AND USE THEREOF

A method is provided for uniformly differentiating a pluripotent stem cell into a neural stem cell, with elimination of variation among cell strains or clones of the pluripotent stem cell and without formation of an embryoid body, regardless of the origin of the pluripotent stem cell 1. A method for uniformly differentiating a pluripotent stem cell into a neural stem cell , with elimination of variation among cell strains or clones of the pluripotent stem cell and without formation of an embryoid body , regardless of the origin of the pluripotent stem cell , comprising:culturing the pluripotent stem cell in a culture medium containing a fibroblast growth factor 2 (FGF 2), a rho-associated protein kinase (ROCK) inhibitor, and a leukemia inhibitory factor (LIF) as active components.2. (canceled)3. The method according to claim 1 , wherein the culturing is carried out under hypoxic conditions.4. The method according to claim 1 , further comprising:dissociating the pluripotent stem cells into individual cells one by one before the culturing.5. The method according to claim 1 , wherein the pluripotent stem cell is a blood cell-derived induced pluripotent stem cell.6. The method according to claim 5 , wherein the induced pluripotent stem cell is a T cell-derived cell.7. A culture medium containing FGF2 claim 1 , a ROCK inhibitor claim 1 , and LIF as active components claim 1 , for use in the method according to .8. A neural stem cell produced by the method according to .9. The neural stem cell according to claim 8 , which expresses a forebrain marker and a forebrain/midbrain marker.10. The neural stem cell according to claim 8 , which does not substantially express homeobox B4 (HOXB 4).11. The neural stem cell according to claim 8 , wherein a T cell receptor gene is rearranged.12. A method for improving the efficiency of differentiating a pluripotent stem cell into a neural stem cell claim 8 , the method comprising:culturing the pluripotent stem cell in a culture medium ...

SERUM-FREE CELL CULTURE MEDIUM

The specification describes an improved serum-free animal cell culture medium, which can used for the production of a protein of interest. Ornithine, or a combination of ornithine and putrescine can be added to serum-free media or chemically defined media to improve viable cell density, to reduce cell doubling time, and to increase the production of a protein of interest. 1. A method for cultivating a cell expressing aflibercept , comprising:(a) providing a cell culture medium comprising 0.6±0.09 mM ornithine and 0.714±0.11 mM putrescine, wherein said medium is serum-free and hydrolysate-free; and(b) propagating or maintaining the cell in said cell culture medium to form a cell culture.2. The method of claim 1 , wherein the cell culture medium is chemically defined.3. The method of claim 1 , wherein the cell culture medium comprises ≥40±6 mM of a mixture of amino acids or salts thereof.4. The method of claim 3 , wherein the mixture of amino acids or salts thereof comprises alanine claim 3 , arginine claim 3 , asparagine claim 3 , aspartic acid claim 3 , cysteine claim 3 , glutamic acid claim 3 , glycine claim 3 , histidine claim 3 , isoleucine claim 3 , leucine claim 3 , lysine claim 3 , methionine claim 3 , phenylalanine claim 3 , proline claim 3 , serine claim 3 , threonine claim 3 , tryptophan claim 3 , tyrosine claim 3 , and valine.5. The method of claim 1 , wherein the cell culture medium comprises one or more fatty acids.6. The method of claim 5 , wherein the one or more fatty acids are selected from the group consisting of linoleic acid claim 5 , thioctic acid claim 5 , oleic acid claim 5 , palmitic acid claim 5 , stearic acid claim 5 , arachidic acid claim 5 , arachidonic acid claim 5 , lauric acid claim 5 , behenic acid claim 5 , decanoic acid claim 5 , dodecanoic acid claim 5 , hexanoic acid claim 5 , lignoceric acid claim 5 , myristic acid and octanoic acid.7. The method of claim 1 , wherein the cell culture medium comprises a mixture of nucleosides.8. ...

METHODS OF EXPANDING EMBRYONIC STEM CELLS IN A SUSPENSION CULTURE

A method of expanding and maintaining human embryonic stem cells (ESCs) in an undifferentiated state by culturing the ESCs in a suspension culture under culturing conditions devoid of substrate adherence is provided. Also provided are a method of deriving ESC lines in the suspension culture and methods of generating lineage-specific cells from ESCs which were expanded in the suspension culture of the present invention. 1. A method of isolating lineage-specific cells from human pluripotent stem cells , comprising:(a) culturing the human pluripotent stem cells in a suspension culture without substrate adherence in the presence of a culture medium which allow expansion of the human pluripotent stem cells in the undifferentiated state for at least 5 passages, thereby obtaining expanded human pluripotent stem cells in an undifferentiated state,(b) differentiating said expanded human pluripotent stem cells in said undifferentiated state into embryoid bodies, and(c) isolating lineage specific cells from said embryoid bodies.2. The method of claim 1 , wherein said culturing without said substrate adherence is effected by culturing the human pluripotent stem cells in a culture vessel having an internal surface designed to prevent attachment or adherence of said pluripotent stem cells to said surface.3. The method of claim 1 , wherein said substrate comprises components of extracellular matrix claim 1 , a glass microcarrier or beads.4. The method of claim 1 , wherein said differentiating is effected by removing said culture medium from said expanded undifferentiated human pluripotent stem cells.5. The method of claim 1 , wherein said isolating is effected by sorting of cells of said embryoid bodies using a fluorescence activated cell sorter (FACS).6. The method of claim 5 , further comprising disaggregating said embryoid bodies prior to incubating cells of said embryoid bodies with a fluorescently-labeled antibody directed against a cell surface antigen characteristics to ...

PROCESS FOR CONTINUOUS CELL CULTURE OF CANCER CELLS AND CANCER STEM CELLS

The present invention is directed towards compositions and methods of culturing cancer cells, with the methods comprising culturing cancer cells in the presence a cell culture medium while inhibiting the activity of Rho kinase (ROCK) in the cells during culturing. 126-. (canceled)27. A composition comprising fibroblast growth factor (FGF) , epithelial growth factor (EGF) , insulin growth factor-1 (IGF-1) , insulin , progesterone , transferrin , putrescine , pyruvate , albumin , selenite , thiamine , glutathione , ascorbic acid , and at least one Rho kinase (ROCK) inhibitor.28. The composition of claim 27 , wherein the composition further comprises glucose.29. The composition of claim 28 , wherein the composition further comprises at least one amino acid.30. The composition of claim 29 , wherein the composition comprises at least one amino acid selected from the group consisting of glycine claim 29 , histidine claim 29 , isoleucine claim 29 , methionine claim 29 , phenylalanine claim 29 , praline claim 29 , hydroxyproline claim 29 , serine claim 29 , threonine claim 29 , tryptophan claim 29 , tyrosine and valine.31. The composition of claim 27 , wherein the composition does not comprise animal serum.32. The composition of claim 27 , wherein the composition further comprises a base cell culture medium.33. The composition of claim 27 , wherein the ROCK inhibitor is an inhibitor of Rho kinase inhibitor 1 (ROCK 1) claim 27 , Rho kinase inhibitor 2 (ROCK 2) or both.34. The composition of claim 33 , wherein the ROCK inhibitor is selected from the group consisting of Y-27632 claim 33 , HA1100 claim 33 , HA1077 claim 33 , Thiazovivin claim 33 , and GSK429286.35. The composition of claim 33 , wherein the ROCK inhibitor is an RNA interference (RNAi) molecule specific for ROCK 1 claim 33 , ROCK 2 or both.36. A cell culture system comprising a composition according to and a culture vessel.37. The cell culture system of claim 36 , wherein the culture vessel comprises ...

METHOD OF DIFFERENTIATING HAIR FOLLICLE CELL INTO GERMLINE STEM CELL AND USE THEREOF

The present invention relates to a method for differentiating hair follicle cells into germline stem cells, germline stem cells differentiated by the method, and use of the same germline stem cells. A method of differentiating hair follicle cells into germline stem cells according to the present invention can differentiate hair follicle cells into germline stem cells using culture conditions only, without genetic modification. Capable of inducing differentiation of cells of specific individual types, such as hair follicle cells, into cells of different types such as germline cells, the present invention is therefore expected to be usefully used for the understanding of reproductive biology and the clinical application thereof. 1. A method for differentiating hair follicle cells into germline stem cells , the method comprising: a step of culturing hair follicle cells on feeder cells using a spermatogonial stem cell differentiation medium containing a growth factor.2. The method of claim 1 , wherein the growth factor is a glial cell line-derived neurotrophic factor(GDNF) claim 1 , GDNF family receptor al(GFRα1) claim 1 , and a basic fibroblast growth factor(bFGF).3. The method of claim 1 , wherein the spermatogonial stem cell differentiation medium is a mouse serum-free medium(mSFM).4. The method of claim 1 , wherein the feeder cells are STO cells.5. The method of claim 1 , wherein the hair follicle cells are cultured for 3 to 5 weeks.6. Germline stem cells differentiated by the method of . The present invention relates to a method for differentiating hair follicle cells into germline stem cells, germline stem cells differentiated by the method, and use of the same germline stem cells.Spermatogenesis is the process during which male gametes or spermatozoa mature, and spermatozoa are produced throughout the entire male lifetime. Spermatogenesis depends on spermatogonial stem cells (SSCs), which are adult stem cells capable of both self-renewal and differentiation. The ...

Mammalian cell culture

The invention provides a method for culturing mammalian cells. The method provides greater control over cell growth to achieve high product titer cell cultures by changing the temperature of the cell culture and/or by starving the cells in their asparagine supply.

Methods for modulating the glycosylation profile of recombinant proteins using sugars

The present invention relates to the field of protein production, and in particular to methods and compositions for modulating glycosylation of recombinant proteins expressed in host cells.

Cell culture improvements

The invention describes improved methods and compositions for producing a recombinant protein, e.g., an antibody, in mammalian cell culture. In addition, the invention provides improved cell culture media, including improved production media, feed solutions, and combination feeds, which may be used to improve protein productivity in mammalian cell culture.

COMPOUNDED MEDIA POWDER FORMULATION AND METHOD OF PREPARATION OF LIQUID MEDIUM FOR CELL CULTURE

The present invention provides a compounded cell culture medium powder formulation comprising: a basal medium powder and a cell culture media supplement, wherein the cell culture media supplement comprises and one or more salts; one or more growth factors; one or more inorganic ions; an amino acid supplement comprising one or more of asparagine, glutamine, histidine, and serine; one or more buffers; and one or more anti-foaming agents. The invention further provides methods of making a compounded cell culture medium powder formulation methods of making a cell culture medium for growing mammalian cells and methods of producing a protein of interest by culturing cells in the cell culture medium and isolating the protein of interest. 1. A compounded cell culture medium powder formulation comprising a basal medium powder and a cell culture media supplement , wherein the cell culture media supplement comprisesi) one or more salts;ii) one or more growth factors;iii) one or more inorganic ions;iv) an amino acid supplement comprising one or more of asparagine, glutamine, histidine, and serine;v) one or more buffers; andvi) one or more anti-foaming agents.2. The compounded cell culture medium powder formulation of claim 1 , wherein the basal medium powder comprises Dulbecco's Modified Eagle's Medium claim 1 , Ham's Medium F12 claim 1 , Eagle's Minimal Essential Medium claim 1 , RPMI 1640 Medium claim 1 , and Dulbecco's Modified Eagle's Medium/Ham's F12 Medium (DMEM/F-12; 1:1 ratio).3. The compounded cell culture medium powder formulation of claim 1 , wherein the basal medium powder comprises one or more of the following components or a combination thereof: biotin claim 1 , calcium chloride claim 1 , choline chloride claim 1 , cyanocobalamin (B12) claim 1 , D+ mannose claim 1 , D-calcium pantothenate claim 1 , dextrose (anhydrous) claim 1 , DL-alpha-lipoic acid claim 1 , ferric nitrate 9HO claim 1 , ferrous sulfate 7HO claim 1 , folic acid claim 1 , glycine claim 1 , ...

METHOD FOR PRODUCING ENGINEERED HEART MUSCLE (EHM)

The present invention provides a new method for producing Engineered Heart Muscle (EHM) under chemically fully defined conditions and compounds all compatible with GMP regulations. The resulting human myocardium generates force and shows typical heart muscle properties. 115-. (canceled)16. A method for producing engineered heart muscle (EHM) , the method comprising the steps of:(i) providing a serum-free reconstitution mixture in one or more moulds, said reconstitution mixture comprising (a) a serum-free minimum essential medium; (b) a serum-free supplement resulting in a final concentration of 0.5-50 mg/ml albumin, 1-100 μg/ml transferrin, 0.1-10 μg/ml ethanol amine, 0.003-0.3 μg/ml sodium selenite, 0.4-40 μg/ml L-Carnitine HCl, 0.1-10 μg/ml Hydrocortisone, 0.05-5 μl/ml Fatty acid supplement, 0.0001-0.1 μg/ml triodo-L-thyronine (T3) and 0.2-2 mg/ml collagen; and (c) a mixture of human cardiac myocytes and human non-myocytes, wherein 20 to 80% of the total cell mixture are cardiac myocytes; wherein the reconstitution mixture has a pH of 7.2 to 7.6;(ii) culturing the serum-free reconstitution mixture in said one or more moulds, whereby the serum-free reconstitution mixture is allowed to condense for at least 15 min;{'sup': '2+', '(iii) culturing the mixture obtained in step (ii) in said one or more moulds in a serum-free EHM culture medium until the mixture condenses to at least 50% of its original thickness, wherein said EHM culture medium comprises (a) a basal medium comprising 0.5-3 mmol/L Ca; (b) a serum-free supplement as defined in (i)(b); (c) 0.5-10 mmol/L L-glutamine; (d) 0.01-1.0 mmol/L ascorbic acid; (e) 1-100 ng/ml IGF-1; and (f) 1-10 ng/ml TGFβ1;'}(iv) culturing the mixture obtained in step (iii) under mechanical stretching in a serum-free EHM culture medium as defined in step (iii) (a)-(f), whereby force-generating EHM is formed.17. The method of claim 16 , wherein the minimum essential medium in step (i) claim 16 , in step (iii) claim 16 , or both in ...

METHOD FOR PRODUCING ADENOHYPOPHYSIS OR PRECURSOR TISSUE THEREOF

The present invention provides a method for inducing adenohypophysis or precursor tissue thereof in vitro from human pluripotent stem cells. The method includes culturing an aggregate of human pluripotent stem cells in suspension in a medium containing a bone morphogenetic protein signal transduction pathway activating substance and a substance acting on the Shh signal pathway to obtain a human cell aggregate containing a hypothalamus neuroepithelial tissue and a surface ectoderm, and further culturing the obtained human cell aggregate containing the hypothalamus neuroepithelial tissue and the surface ectoderm in suspension in a medium containing a bone morphogenetic protein signal transduction pathway activating substance and a substance acting on the Shh signal pathway to induce formation of hypophysial placode and/or Rathke's pouch in the surface ectoderm, thus obtaining a human cell aggregate containing 1) hypothalamus neuroepithelial tissue, and 2) hypophysial placode and/or Rathke's pouch. 1. A method for producing a human cell aggregate comprising adenohypophysis or a precursor tissue thereof , which comprises culturing an aggregate of human pluripotent stem cells in suspension in a medium containing a bone morphogenetic protein signal transduction pathway activating substance and a substance acting on the Shh signal pathway.2. The production method according to claim 1 , comprising culturing an aggregate of human pluripotent stem cells in suspension in a medium containing a bone morphogenetic protein signal transduction pathway activating substance and a substance acting on the Shh signal pathway to obtain a human cell aggregate comprising a hypothalamus neuroepithelial tissue and a surface ectoderm by claim 1 , and further culturing the obtained human cell aggregate comprising the hypothalamus neuroepithelial tissue and the surface ectoderm in suspension in a medium containing a bone morphogenetic protein signal transduction pathway activating substance and ...

Media for Cell Culture

The present disclosure relates, in general, to a media, e.g., a serum replacement, media supplement, complete media or cryopreservation media, comprising a base physiological buffer and liposomes comprising cholesterol, phosphatidylcholine and fatty acids. It is contemplated that media provides advantages to improve cell growth in culture compared to cells cultured not using the serum replacement described herein.

CULTURE MEDIUM AND CULTURING METHOD FOR ANCHORAGE-DEPENDENT CELLS, CELL COMPOSITION INCLUDING STEM CELLS AND/OR DIFFERENTIATED CELLS DERIVED FROM STEM CELLS, AND PRODUCTION METHOD FOR CELL COMPOSITION

As a technique capable of culturing anchorage-dependent cells without using an anchorage, provided is a medium for anchorage-dependent cells, which comprises MFG-E8 (Milk fat globule-EGF factor 8) or a fragment of the protein. This medium can promote adhesion of anchorage-dependent cells in the absence of an anchorage, enables the survival and proliferation (colony formation) of the cells, and further, also enables the subsequent differentiation if the anchorage-dependent cells are stem cells. 1. A medium for an anchorage-dependent cell , which comprises MFG-E8 (Milk fat globule-EGF factor 8) or a fragment of the protein.2. The medium according to claim 1 , wherein the anchorage-dependent cell is a stem cell.3. The medium according to claim 2 , wherein the stem cell is an embryonic stem cell claim 2 , an induced pluripotent stem cell claim 2 , or a mesenchymal stem cell.4. The medium according to claim 1 , wherein the anchorage-dependent cell is derived from a human.5. The medium according to claim 1 , which is a serum-free medium.6. An adhesion promoter for an anchorage-dependent cell claim 1 , which comprises MFG-E8 or a fragment of the protein.7. A method for culturing an anchorage-dependent cell claim 1 , which comprises a step of culturing an anchorage-dependent cell in a medium comprising MFG-E8 or a fragment of the protein in the absence of an anchorage.8. The culture method according to claim 7 , wherein the anchorage-dependent cell is a stem cell.9. The culture method according to claim 8 , wherein the stem cell is an embryonic stem cell claim 8 , an induced pluripotent stem cell claim 8 , or a mesenchymal stem cell.10. The culture method according to claim 7 , wherein the anchorage-dependent cell is derived from a human.11. The culture method according to claim 7 , wherein the medium is a serum-free medium.12. A method for producing a cell composition comprising a stem cell claim 7 , wherein the method comprises a step of culturing a stem cell in a medium ...

COMPOSITIONS CONTAINING PLATELET CONTENTS

This document provides methods and materials relating to platelet lysates. For example, methods and materials for using platelet lysate compositions to grow adult stem cells, to differentiate adult stem cells, to grow primary cell cultures, to grow tumor stem cells, and to identify effective growth factors are provided. 118-. (canceled)19. A stable human platelet lysate composition comprising: i) plasma,', 'ii) a total protein concentration of at least about 35 mg per mL,', 'iii) a protein fraction that contains growth factor activity;', 'iv) greater than 200 pg human VEGF per mL; and, 'a lysed platelet preparation comprisingwherein said platelet lysate composition comprises protein complexes greater than 50 kDa.21. The platelet lysate composition of claim 19 , wherein the composition further comprises: about 180 pg/mL FGF.22. The platelet lysate composition of claim 19 , wherein the composition further comprises: about 54 ng/mL TGF-β.23. The platelet lysate composition of claim 19 , wherein the composition further comprises: about 8.5 ng/mL PDGF-BB.24. The platelet lysate composition of claim 19 , wherein the composition further comprises: about 180 pg/mL FGF; about 8.5 ng/mL PDGF; about 132 ng/mL IGF-1; and about 54 ng/mL TGF-β.24. The platelet lysate composition of claim 19 , wherein the total protein concentration is least about 48 mg per mL.25. The platelet lysate composition of claim 19 , wherein the platelet lysate composition is passed through a 0.20 μm or smaller filter after begin filtered by a 0.45 μm or smaller filter.26. The platelet lysate composition of claim 19 , wherein the composition further comprises heparin.27. The platelet lysate composition of claim 19 , wherein the composition is a spray claim 19 , glue claim 19 , or a suture.28. The platelet lysate composition of claim 26 , wherein the composition is a spray claim 26 , glue claim 26 , or a suture.29. The platelet lysate composition of claim 19 , wherein the protein fraction has a molecular ...

CULTURE MEDIUM AND METHOD FOR ENRICHING AND MAINTAINING CANCER STEM CELLS (CSCS) USING SAID MEDIUM

The present invention relates to a serum-free conditioned medium that solves the drawbacks mentioned in the prior art, as it does not require prior handling of the cells, and it furthermore allows starting from a large population with no additional cost. This medium favors in vitro proliferation and conservation of the pluripotency potential that allows maintaining a state that is undifferentiated with respect to the subpopulation of cancer stem cells (CSCs) and in turn does not allow survival of the differentiated cells. 1. A cell culture medium suitable for isolating and/or enriching cancer stem cells (CSCs) , obtained or obtainable by a method comprising the following steps:a. Seeding mesenchymal stem cells (MSCs) in a plate, vessel or flask suitable for culturing cells using a culture medium suitable for this purpose;b. Optionally removing any remainder of medium and/or serum from the culture medium from step a) by means of washing;c. Adding a sphere medium or conventional medium without FBS to the product of step b);d. Collecting the culture medium from step c) and adding new sphere medium;e. Repeating the process from step d) until the cells in culture reach a level of confluence of 80-90%;f. Optionally filtering and freezing the medium obtained in steps d) and e) until use.2. The cell culture medium according to claim 1 , wherein said medium is obtained or obtainable by a method comprising the following steps:a. Seeding mesenchymal stem cells in a plate, vessel or flask suitable for culturing cells using a conventional culture medium, where said cells have a confluence between 40% and 90% at the start of the process;b. Removing any remainder of medium and/or serum from the culture medium from step a) by means of washing at least 24 hours after the start of the process;c. Adding a sphere medium or conventional medium without FBS to the product of step b);d. Collecting the culture medium from step c) at least 48 hours after adding the medium in step c), and ...

COMPOSITIONS AND METHODS TO DERIVE MESODERMAL LINEAGE CELLS AND MIXED TISSUE ORGANOIDS FROM EMBRYONIC STEM CELLS

The disclosure relates to methods for producing mesodermal lineage cells, cardiac lineage cells, hematopoietic lineage cells, retinal lineage cells, and combinations thereof. In some aspects, the disclosure relates to methods of producing mixed tissue organoids comprising retinal lineage cells and cardiac lineage cells. When interaction of WDR5 protein at the binding pocket/interaction surface with RBBP5/MYC/KANSL2 is disrupted in the embryonic stem cell for a set period of time after differentiation of the embryonic stem cell has begun, and the disruption of that interaction is subsequently removed, differentiation of the embryonic stem cell to a mesodermal lineage cell is obtained. Such mesodermal lineage cells may, in some methods of the disclosure, be subsequently differentiated into cardiac lineage cells and hematopoietic lineage cells. The disclosure also relates to method for producing mixed lineage organoids, comprising retinal and mesodermal, including cardiac, lineage cells in a single organoid. The disclosure also relates to cells and organoids obtained by the methods, uses of the cells and organoids, and kits comprising the cells and/or reagents for producing them. 1. A method for producing in culture a mesodermal lineage cell from an embryonic stem cell comprising recombinant WD repeat domain 5 (WDR5) protein , the method comprising:a) disrupting interaction of recombinant WDR5 protein with retinoblastoma-binding protein 5 (RBBP5), MYC, or KATE Regulatory NSL Complex Subunit 2 (KANSL2) protein in the embryonic stem cell for a set period of time, wherein the set period of time is greater than 24 hours and less than 60 hours, andb) removing the disruption of the interaction of recombinant WDR5 protein with RBBP5, MYC, or KANSL2 in the embryonic stem cell to allow the mesodermal lineage cell to differentiate from the embryonic stem cell.2. The method of claim 1 , wherein the disrupting step is carried out by silencing recombinant WDR5 protein expression by ...

Cell Culture Media Composition and Methods of Producing Thereof

A serum free cell culture media, wherein the media is adapted to be conditioned by culturing a first set of eukaryotic cells in the media, wherein the first set of eukaryotic cells use an expression vector to excrete levels of desired complex proteins into the media; wherein said desired complex proteins include human Growth Hormone (hGH), Growth Hormone-like growth factors, insulin-like growth factors, insulin, modified insulins, cytokines, mitogenic proteases and mixtures thereof; and wherein the media is adapted to grow a set of eukaryotic cells. 120.-. (canceled)21. A co-culture of cells comprising , a Chinese Hamster Ovary (CHO) feeder cell line comprising an expression vector encoding human Growth Hormone (hGH) and a second set of mammalian cells , wherein the CHO feeder cell line secretes hGH into the co-culture and promotes transfection of the second set of mammalian cells during co-culture.22. A co-culture of cells according to claim 21 , wherein the second set of mammalian cells are CHO cells.23. A co-culture of cells according to claim 21 , wherein the second set of mammalian cells is selected from the group consisting of CHO-K1 claim 21 , CHO-DG DHFR- and CHO-S.24. A co-culture of cells according to claim 21 , wherein the cells in co-culture are seeded in single wells of a microtitre plate comprising cell culture media.25. A co-culture of cells comprising claim 21 , a Chinese Hamster Ovary (CHO) feeder cell line comprising an expression vector encoding human growth hormone (hGH) and a second set of mammalian cells that produce a desired recombinant protein claim 21 , wherein the CHO feeder cell line secretes hGH into the co-culture.26. A co-culture of cells according to claim 25 , wherein the second set of mammalian cells are CHO cells.27. A co-culture of cells according to claim 25 , wherein the second set of mammalian cells is selected from the group consisting of CHO-K1 claim 25 , CHO-DG DHFR- and CHO-S.28. A co-culture of cells according to claim 25 ...

CELL CULTURE IMPROVEMENTS

The invention describes improved methods and compositions for producing a recombinant protein, e.g., an antibody, in mammalian cell culture. In addition, the invention provides improved cell culture media, including improved production media, feed solutions, and combination feeds, which may be used to improve protein productivity in mammalian cell culture. 1. A serum-free cell culture medium comprising Part A , Part B , and Part C , whereina) Part A consists essentially of a modified basal medium which excludes the following components: sodium bicarbonate, a buffer, mono-basic sodium phosphate, di-basic sodium phosphate, an osmolarity regulator, a surfactant, and monosaccharide glucose;b) Part B consists essentially of an inorganic iron source; andc) Part C comprises a recombinant growth factor; a buffer; an osmolarity regulator; an energy source; and at least two different non-animal hydrolysates.234-. (canceled)35. A serum-free cell culture medium comprising:a) a basal medium;b) about 8-12 ml/kg or 116-126 mg/L ferric citrate;c) about 2-6 mg/kg recombinant human insulin;d) about 2-5 g/kg anhydrous glucose;e) about 0.1-0.5 g/kg L-glutamine;f) about 1-3 g/kg sodium bicarbonate;{'sub': 2', '4', '2, 'g) about 0.01-0.05 g/kg NaHPO.HO;'}{'sub': 2', '4', '2, 'h) about 0.4 to 0.5 g/kg of NaHPO.7HO; and'}i) about 1.0-3.0 g/kg yeast-based hydrolysate.36. The cell culture medium of claim 35 , comprisinga) a basal medium;b) about 10.0 ml/kg or 122 mg/L ferric citrate;c) about 4.0 mg/kg recombinant human insulin;d) about 3.5 g/kg anhydrous glucose;e) about 0.29 g/kg L-glutamine;f) about 1.6 g/kg sodium bicarbonate;{'sub': 2', '4', '2, 'g) about 0.03 g/kg NaHPO.HO;'}{'sub': 2', '4', '2, 'h) about 0.43 to 0.44 g/kg of NaHPO.7HO; and'}i) about 2.0 g/kg yeast-based hydrolysate.3741-. (canceled)42. A serum-free cell culture production medium comprising:a) a modified basal medium which excludes the following components sodium bicarbonate, buffer, mono-basic sodium phosphate, di- ...

CELL CULTURE MEDIUM FOR EUKARYOTIC CELLS

Cell culture media are provided herein as are methods of using the media for cell culture and protein production from cells. 1. A method for culturing eukaryotic cells for increasing production of a protein , comprising the steps of:culturing cells in a defined cell culture medium;supplementing the cell culture medium with nicotinamide, wherein concentration of the nicotinamide is about 50 nM to about 2000 nM; andproducing a protein in the eukaryotic cells,wherein the supplementation with nicotinamide increases a titer of the protein.2. The method of claim 1 , wherein the titer of the protein is at least about 2% greater than another method with a cell culture medium that does not have at least about 50 nM nicotinamide.3. The method of claim 1 , wherein the eukaryotic cells include at least one selected from the group consisting of: Baby Hamster Kidney cell lines claim 1 , Chinese Hamster Ovary cell lines claim 1 , Murine myeloma cell lines claim 1 , Mouse myeloma cell lines claim 1 , Human embryonic kidney cell lines claim 1 , Human-retina-derived cell lines claim 1 , and Amniocyte cell lines.4. The method of claim 1 , wherein the protein is secreted in the medium.5. The method of claim 1 , wherein the cell culture medium does not have a protein derived from an animal.6. The method of claim 1 , wherein the cell culture medium is a serum-free medium.7. The method of claim 1 , wherein the cell culture medium is a chemically-defined medium.8. A method for producing a protein claim 1 , comprising:introducing into a cell a nucleic acid comprising a nucleotide sequence encoding a protein;culturing the cell in a cell culture medium comprising at least about 50 nM nicotinamide or at least about 10 nM 5-methythioadenosine; andproducing the protein in the cell. The present invention generally pertains to a cell culture medium for eukaryotic cells and the production of natural and recombinant products derived therefrom.Cell culture manufacturing technology is widely used for ...

METHODS OF GENERATING T-CELLS FROM STEM CELLS AND IMMUNOTHERAPEUTIC METHODS USING THE T-CELLS

Methods and composition for production of T cells are provided. Also provided are therapeutic methods using engineered T cells. For example, in certain aspects methods include preparing three dimensional cell culture compositions comprising stroma cells and hematopoietic stem or progenitor cells in a serum-free medium for producing T cells. 1. (canceled)3. The method of claim 2 , wherein the Notch ligand is an exogenous Notch ligand.4. The method of claim 2 , wherein the method further comprises centrifugation of the stem or progenitor cells and the stromal cells to form a 3D cell aggregate.5. The method of claim 2 , wherein the medium further comprises externally added ascorbic acid.6. The method of claim 2 , wherein the medium further comprises externally added FLT3 ligand (FLT3L) claim 2 , interleukin 7 (IL-7) claim 2 , stem cell factor (SCF) claim 2 , thrombopoietin (TPO) claim 2 , stem cell factor (SCF) claim 2 , thrombopoietin (TPO) claim 2 , IL-2 claim 2 , IL-4 claim 2 , IL-6 claim 2 , IL-15 claim 2 , IL-21 claim 2 , TNF-alpha claim 2 , TGF-beta claim 2 , interferon-gamma claim 2 , interferon-lambda claim 2 , TSLP claim 2 , thymopentin claim 2 , pleotrophin claim 2 , midkine claim 2 , or combinations thereof.7. The method of claim 2 , wherein the medium further comprises one or more vitamins claim 2 , proteins claim 2 , amino acids claim 2 , monosaccharides claim 2 , inorganic ions claim 2 , molybdenum claim 2 , vanadium claim 2 , iron claim 2 , zinc claim 2 , selenium claim 2 , copper claim 2 , manganese claim 2 , corticosterone claim 2 , D-Galactose claim 2 , ethanolamine claim 2 , glutathione claim 2 , L-carnitine claim 2 , linoleic acid claim 2 , linolenic acid claim 2 , progesterone claim 2 , putrescine claim 2 , sodium selenite claim 2 , and triodo-I-thyronine.819-. (canceled)20. The method of claim 2 , wherein the stromal cells have an exogenous nucleotide sequence encoding a Notch ligand claim 2 , and wherein the Notch ligand comprises intact claim 2 ...

Method of Enhancing Cell Growth Using Alkyl-Amine-N-Oxide (AANOX)

The present invention relates to a method to enhance cell growth in culture comprising adding an alkyl-amine-n-oxide (AANOx), such as dodecyldimethylamine oxide (DDAO), into the culture medium in an amount sufficient to improve cell growth. 119-. (canceled)20. A cell culture media comprising alkyl-amine-n-oxide (AANOx) in an amount sufficient to enhance cell growth when used in the culture media.21. The media of wherein the AANOx is dodecyldimethylamine oxide (DDAO) or a related analyte.22. The media of wherein the amount of AANOx is between about 4 and about 80 ppb.23. The media of wherein the amount of AANOx is between about 4 and about 50 ppb.24. The media of wherein the amount of AANOx is between about 10 ppb and about 40 ppb.25. The media of wherein the AANOx is not derived from a soy hydrolysate preparation.26. The media of wherein the AANOx is derived from a soy hydrolysate preparation.27. The media of claim 20 , wherein the AANOx is a DDAO-related analyte having an alkyl chain selected from the group consisting of a C10 claim 20 , C12 claim 20 , C14 claim 20 , and C16 alkyl.28. The media of claim 27 , wherein the DDAO-related analyte is selected from the group consisting of dimethyl-tetradecyl-amine-oxide or dimethyl-hexadecyl-amine-oxide.29. The media of claim 20 , wherein the culture media is animal protein-free media.30. The media of claim 20 , wherein the culture media comprises animal protein.31. The media of wherein the cells are mammalian cells.32. The media of claim 20 , wherein the cells are selected from the group consisting of BSC cells claim 20 , LLC-MK cells claim 20 , CV-1 cells claim 20 , COS cells claim 20 , VERO cells claim 20 , MDBK cells claim 20 , MDCK cells claim 20 , CRFK cells claim 20 , RAF cells claim 20 , RK cells claim 20 , TCMK-1 cells claim 20 , LLCPK cells claim 20 , PK15 cells claim 20 , LLC-RK cells claim 20 , MDOK cells claim 20 , BHK-21 cells claim 20 , CHO cells claim 20 , NS-1 cells claim 20 , MRC-5 cells claim 20 , WI-38 ...

Methods of expanding embryonic stem cells in a suspension culture

A method of expanding and maintaining human embryonic stem cells (ESCs) in an undifferentiated state by culturing the ESCs in a suspension culture under culturing conditions devoid of substrate adherence is provided. Also provided are a method of deriving ESC lines in the suspension culture and methods of generating lineage-specific cells from ESCs which were expanded in the suspension culture of the present invention.

METHOD FOR PRODUCING ENGINEERED HEART MUSCLE (EHM)

The present invention provides a new method for producing Engineered Heart Muscle (EHM) under chemically fully defined conditions all compatible with GMP regulations. The resulting human myocardium generates force and shows typical heart muscle properties. 1. A method for producing engineered heart muscle (EHM) , the method comprising the steps of:(i) providing a serum-free reconstitution mixture in one or more moulds, said reconstitution mixture comprising (a) a serum-free minimum essential medium; (b) a serum-free supplement resulting in a final concentration of 0.5-50 mg/ml albumin, 1-100 μg/ml transferrin, 0.1-10 μg/ml ethanol amine, 0.003-0.3 μg/ml sodium selenite, 0.4-40 μg/ml L-Carnitine HCl, 0.1-10 μg/ml Hydrocortisone, 0.05-5 μl/ml Fatty acid supplement, 0.0001-0.1 μg/ml triodo-L-thyronine (T3) and 0.2-2 mg/ml collagen;and (c) a mixture of human cardiac myocytes and human non-myocytes, wherein 20 to 80% of the total cell mixture are cardiac myocytes;wherein the reconstitution mixture has a pH of 7.2 to 7.6;(ii) culturing the serum-free reconstitution mixture in said one or more moulds, whereby the serum-free reconstitution mixture is allowed to condense for at least 15 min;{'sup': '2+', '(iii) culturing the mixture obtained in step (ii) in said one or more moulds in a serum-free EHM culture medium until the mixture condenses to at least 50% of its original thickness, wherein said EHM culture medium comprises (a) a basal medium comprising 0.5-3 mmol/L Ca; (b) a serum-free supplement as defined in (i)(b); (c) 0.5-10 mmol/L L-glutamine; (d) 0.01-1.0 mmol/L ascorbic acid; (e) 1-100 ng/ml IGF-1; and (f) 1-10 ng/ml TGFβ1; and'}(iv) culturing the mixture obtained in step (iii) under mechanical stretching in a serum-free EHM culture medium as defined in step (iii) (a)-(f), whereby force-generating EHM is formed.2. The method of claim 1 , wherein the minimum essential medium in step (i) claim 1 , in step (iii) claim 1 , or both in step (i) and step (iii) is selected ...

METHODS FOR TREATING MYELOID MALIGNANCIES

The invention relates to methods of treating myeloid malignancies by administering compositions comprising Vδ1+T cells. 1. A method of treating a myeloid malignancy comprising administering a therapeutically effective amount of an allogeneic composition comprising Vδ1+ T cells to a patient with said myeloid malignancy.2. The method as defined in claim 1 , wherein the myeloid malignancy is selected from acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS).3. The method as defined in claim 1 , wherein the patient is positive for minimal residual disease (MRD+).4. The method as defined in claim 3 , wherein the MRD+ patient is in complete remission claim 3 , contains no detectable leukemic blasts in the peripheral blood and contains less than 5% leukemic blasts in the bone marrow.5. The method as defined in claim 1 , which additionally comprises administration of chemotherapy.6. The method as defined in claim 5 , wherein the patient is treated with chemotherapy at least 3 days prior to administration of the allogeneic composition.7. The method as defined in claim 5 , wherein the chemotherapy is selected from fludarabine and cyclophosphamide.8. The method as defined in claim 1 , wherein the therapeutically effective amount comprises about 8×10 claim 1 , 4×10 claim 1 , 2.4×10 claim 1 , 1.2×10 claim 1 , 8×10 claim 1 , 4×10 claim 1 , 8×10or 4×10total live cells.9. The method as defined in claim 1 , wherein the therapeutically effective amount comprises less than about 1×10total live cells.10. The method as defined in claim 1 , wherein the therapeutically effective amount comprises less than about 1×10total live cells.11. The method as defined in claim 1 , wherein the therapeutically effective amount comprises less than about 1×10total live cells.12. The method as defined in claim 8 , wherein the therapeutically effective amount comprises at least about 90% CD45+ cells relative to total live cells.13. The method as defined in claim 8 , wherein the therapeutically ...

METHODS OF DIFFERENTIATION TO NEURONAL CELLS AND KITS THEREFOR

Embodiments herein provide methods of differentiating neural stem cells to neuronal cells while concomitantly retarding neural stem cell proliferation. Resultant cultures demonstrate reduced clumping of cells, increased purity of neuronal cells and accelerated electrophysiology as compared to control methods. 1. A method for accelerating differentiation of at least one neural stem cell to at least one neuronal cell and concomitantly retarding neural stem cell proliferation , comprising:culturing the at least one neural stem cell in a differentiation medium for a time and under conditions to form the at least one neuronal cell,wherein the differentiation medium comprises a serum-free neural stem cell culture medium, and a serum-free supplement comprising at least one gamma secretase inhibitor,wherein excitability of the at least one neuronal cell is accelerated as compared to culturing the at least one neural stem cell in the differentiation medium lacking the at least one gamma secretase inhibitor.2. The method of wherein excitability of the at least one neuronal cell is accelerated by at least 100% at Day 7 of differentiation.3. The method of claim 1 , wherein the at least one neural stem cell is derived from an induced pluripotent stem cell.4. The method of claim 1 , wherein the at least one neural stem cell is derived from an embryonic stem cell.5. The method of wherein the at least one neural stem cell is a SOX1 positive neural stem cell and the at least one neuronal cell is a MAP2 positive neuronal cell.6. The method of claim 1 , wherein the serum-free supplement of the differentiation medium comprises a gamma secretase inhibitor selected from the group consisting of Compound E claim 1 , YO-01027 claim 1 , LY411575 claim 1 , MK-0752 claim 1 , a salt thereof claim 1 , and a combination thereof.7. The method of claim 6 , wherein the serum-free supplement of the differentiation medium comprises a gamma secretase inhibitor selected from the group consisting of ...

LARGE SCALE CELL CULTURE SYSTEM FOR MAKING MEAT AND ASSOCIATED PRODUCTS

Provided is a large-scale cell culture system for producing products without harming animals. Also provided is a method for making meat products using this cell culture system. Further provided is a method for making personal care products using this cell culture system, as well as a method for making nutritional supplements using this cell culture system. 1. A cell culture system comprising;a pump; at least two culturing vessels; a fresh basal medium vessel; and a waste collecting vessel, wherein at least two culturing vessels, the fresh basal medium vessel and the waste collecting vessel are connected in parallel with means for enabling a serum-free medium to move between the vessels.2. The system of claim 1 , wherein each culturing vessel contains a different type of cells selected from the group consisting of muscle claim 1 , fat claim 1 , cartilage claim 1 , liver claim 1 , heart claim 1 , kidney claim 1 , and lung claim 1 , and other mammalian cells.3. The system of claim 1 , wherein a single culturing vessel contains multiple types of cells wherein the cells are selected from the group consisting of muscle claim 1 , fat claim 1 , cartilage claim 1 , liver claim 1 , heart claim 1 , kidney claim 1 , and lung claim 1 , and other mammalian cells.4. A method for making conditioned medium comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'providing the system of with a serum-free medium;'}adding animal cells into the culturing vessels;circulating the serum-free medium between the vessels;heating the system to a temperature suitable for culturing; andagitating and incubating the cells in the vessels for a time sufficient to culture the cellsto a desired cell density.5. The method of claim 4 , wherein each culturing vessel contains a different type of cells selected from the group consisting of muscle claim 4 , fat claim 4 , cartilage claim 4 , liver claim 4 , heart claim 4 , kidney claim 4 , and lung claim 4 , and other mammalian cells.6. The system of ...

NEW MEDIUM FOR HIGH PERFORMANCE MAMMALIAN FED-BATCH CULTURES

The present invention relates to new serum- and protein-free culture media. These media are high performance culture media, which notably improve mammalian fed-batch cultures. The present invention also relates to methods for preparing and/or designing the medium, and methods of use thereof. 1. A cell culture medium comprising NaHPO , L-Leucine , L-Lysine , L-Methionine , L-Glutamic acid , L-phenylalanine , L-proline , L-threonine , L-tryptophan , L-Valine , magnesium sulfate , calcium chloride , myo-inositol , sodium pyruvate , D-Biotin , choline chloride , L-Aspargine , folic acid , niacinamide (B3) , D-pantothenic acid×½Ca , L-Serine , potassium chloride , pyridoxine , L-Aspartic acid , riboflavin , thiamine , ferric ammonium citrate , vitamin B12 , hypoxanthine , thymidine , putrescine , ethanolamine , zinc sulfate , cupric sulfate , pluronic , L-tyrosine , sodium selenite , L-arginine , L-Cysteine , L-Histidine and L-Isoleucine , wherein the medium is serum and protein free.2. The medium according to comprising: 1.7 to 10 mM of NaHPO claim 1 , 2 to 9 mM of L-Leucine claim 1 , 1 to 6 mM of L-Lysine claim 1 , 0 to 3 mM of Glycine claim 1 , 0.4 to 2 mM of L-Methionine claim 1 , 1 to 4 mM of L-Glutamic acid claim 1 , 0.5 to 3 mM of L-phenylalanine claim 1 , 0.7 to 6 mM of L-proline claim 1 , 0.7 to 6 mM of L-threonine claim 1 , 0.5 to 2 mM of L-tryptophan claim 1 , 1 to 7 mM L-Valine claim 1 , 0.1 to 1.5 mM of Magnesium Sulfate claim 1 , 0.1 to 1.05 mM of Calcium Chloride claim 1 , 0.07 to 0.7 mM of myo-Inositol claim 1 , 0.8 to 4 mM of Sodium pyruvate claim 1 , 0.0008 to 0.01 mM of D-Biotin claim 1 , 0.1 to 1 mM of Choline Chloride claim 1 , 3 to 9 mM of L-Aspargine claim 1 , 0.006 to 0.04 mM of Folic acid claim 1 , 0.03 to 0.15 mM of Niacinamide (B3) claim 1 , 0.015 to 0.15 mM of D-pantothenic acid×½Ca claim 1 , 1 to 8 mM of L-Serine claim 1 , 1 to 10 mM of Potassium Chloride claim 1 , 0.005 to 0.05 mM of Pyridoxine claim 1 , 0.8 to 2.4 mM of L-Aspartic acid ...

CELL CULTURE MEDIA COMPOSITIONS FOR PRIMARY CELLS

Platelet lysate compositions and cell culture media compositions for maintaining and/or growing mammalian cells, such as mammalian endothelial cells (ECs) and mammalian endothelial progenitor cells (EPCs), in particular human ECs (huECs) and human EPCs (huEPCs), such as primary huECs and primary huEPCs, are provided. The cell culture media compositions contain a basal medium, a platelet lysate and, optionally, one or more exogenously added growth factors. Also provided are methods for making and using such cell culture media compositions to grow and/or maintain ECs and EPCs, including huECs and huEPCs, as well as cell culture vessels, dishes, plates, and/or flasks pretreated with the cell culture media compositions. 1. A lyophilized composition comprising a) a human platelet lysate; and b) a physiological carrier; wherein the lyophilized composition does not include a bulking agent.2. The lyophilized composition of claim 1 , wherein the lyophilized composition does not include a bulking agent selected from the group consisting of mannitol claim 1 , sucrose claim 1 , and trehalose.3. The lyophilized composition of claim 1 , wherein the physiological carrier is physiological saline.4. The lyophilized composition of claim 1 , wherein the composition further comprises a supplemental growth factor.5. The lyophilized composition of claim 4 , wherein the supplemental growth factor is FGF-B.6. The lyophilized composition of claim 5 , wherein the supplemental growth factor is present in the composition at a ratio of at least 2000 to 1 or less per ng of FGF in the platelet lysate.7. A solid substrate coated with a lyophilized platelet lysate of .8. (canceled)9. (canceled)10. (canceled)11. (canceled)12. (canceled)13. (canceled)14. (canceled)15. (canceled)16. (canceled)17. (canceled)18. (canceled)19. A wound healing composition comprising a lyophilized platelet lysate having 50% or less of water claim 1 , a supplemental growth factor in a ratio of at least 100 to 1 or less per ...

Serum-Free Chemically Defined Cell Culture Medium

Embodiments of chemically defined cell culture media containing nutrients and growth factors free of any serum for culturing cells such as mesenchymal stem cells and methods of using embodiments of the cell culture medium for expanding cell populations such as mesenchymal stem cells while maintaining a pluripotent phenotype and methods of inducing chondrogenesis and osteogenesis of mesenchymal stem cells by admixing differentiation factors into embodiments of the cell culture medium.

MEDIA FOR CULTURING PLURIPOTENT STEM CELLS

A culture medium is disclosed which comprises STAT3 activator, an ERK1/2 inhibitor and an Axin stabilizer, and optionally also a PKC inhibitor. Cell cultures comprising same and uses thereof are also disclosed. 1. A culture medium comprising a STAT3 activator , an ERK1/2 inhibitor and an Axin stabilizer.2. The culture medium of claim 1 , further comprising a PKC inhibitor.3. A culture medium comprising a STAT3 activator claim 1 , an ERK1/2 inhibitor claim 1 , an Axin stabilizer and a PKC inhibitor.4. The culture medium of claim 1 , further comprising at least one agent selected from the group consisting of: a ROCK inhibitor claim 1 , an SRC inhibitor claim 1 , ascorbic acid claim 1 , a PKA agonist claim 1 , a YAP/TAZ inhibitor claim 1 , a NOTCH inhibitor claim 1 , an SHH inhibitor claim 1 , a TGFβR inhibitor claim 1 , a BMP inhibitor claim 1 , an FGFR inhibitor claim 1 , a JNK inhibitor claim 1 , an ERK5 inhibitor claim 1 , a BRAF inhibitor claim 1 , an ARAFi claim 1 , a CRAFi claim 1 , a p38 inhibitor claim 1 , a GSK3b inhibitor claim 1 , an LSD1 inhibitor claim 1 , a PI3K activator claim 1 , a SMAD activator and a DOT1L inhibitor.5. The culture medium of claim 4 , wherein said SMAD activator is selected from the group consisting of: activin A claim 4 , TGFβ1 and BMP4.6. The culture medium of claim 1 , further comprising a GSK3b inhibitor.7. (canceled)8. The culture medium of claim 1 , further comprising at least one agent selected from the group consisting of: basic fibroblast growth factor (bFGF) claim 1 , a transforming growth factor receptor (TGFR) inhibitor claim 1 , a GSK3b inhibitor claim 1 , a ROCK inhibitor claim 1 , a P38 inhibitor claim 1 , a JNK inhibitor claim 1 , a NOTCH inhibitor claim 1 , a SRC inhibitor claim 1 , insulin-like growth factor 1 (IGF1) claim 1 , insulin-like growth factor II (IGFII) claim 1 , a bone morphogenetic protein (BMP) signaling inhibitor claim 1 , a Sonic Hedgehog pathway (SHH) inhibitor claim 1 , an ERK5 inhibitor claim 1 , ...

METHOD FOR PREPARING INDUCED PLURIPOTENT STEM CELLS USING SYNTHETIC PEPTIDE

Provided is a method of preparing induced pluripotent stem cells using a synthetic peptide, and more particularly, to a method of preparing induced pluripotent stem cells using a peptide capable of inhibiting the activity of NF-κB and promoting mesenchymal-epithelial transition (MET). Since undifferentiated multipotent stem cells may be efficiently prepared under xenopathogen-free or feeder cell-free conditions without requiring co-culture with animal serum or xenogeneic cells, the method for preparing the induced pluripotent stem cells using the synthetic peptide according to the present disclosure is very useful for developing stem cell therapeutic agents that are clinically applicable. 2. The composition of claim 1 , wherein the peptide is any one selected from the group consisting of SEQ ID NOs: 1 to 3.3. The composition of claim 1 , wherein the differentiated cells are somatic cells or precursor cells.4. The composition of claim 1 , wherein the differentiated cells are derived from a human.6. The method of claim 5 , wherein the peptide is any one selected from the group consisting of SEQ ID NOs: 1 to 3.7. The method of claim 5 , wherein the peptide has a concentration of 0.01 to 100 μM.8. The method of claim 5 , wherein the differentiated cells are somatic cells or precursor cells.9. The method of claim 5 , wherein the differentiated cells are derived from a human.10. The method of claim 5 , wherein the induced pluripotent stem cells are prepared under xenopathogen-free or feeder cell-free conditions.11. The method of claim 5 , wherein the reprogramming gene is any one selected from the group consisting of Oct3/4 claim 5 , Sox2 claim 5 , c-Myc claim 5 , Klf4 and Lin28.12. The method of claim 5 , wherein the peptide inhibits NF-κB activity.13. The method of claim 5 , wherein the peptide promotes mesenchymal-epithelial transition (MET).14. The method of claim 5 , wherein the cells to which the reprogramming gene has been introduced are treated by the peptide in a ...

Methods of in vitro Oocyte Development

Methods of preparing ovarian tissue for primordial follicle growth are presented comprising the steps: providing an ovarian tissue sample comprising cortical tissue and stromal tissue; removing damaged tissue from the ovarian tissue sample where present; removing excess stromal tissue from the ovarian tissue sample where present; and then mechanically stretching the ovarian tissue sample along at least one dimension of the ovarian tissue sample, such that the size of the ovarian tissue sample along the at least one dimension is increased by at least 10%. Methods of growing viable oocyte in vitro, and methods of preparing individual ovarian follicles for growth are also presented.

Hematopoietic Stem and Progenitor Cell Expansion System

Described herein is a growth medium for culture of stem cells and/or primary cells, in particular hematopoietic stem cells (HSCs), the growth medium including a basal medium and a supplement, with the medium and/or supplement including a histone acetyltransferase (HAT) inhibitor, a histone deacetylase (HDAC) inhibitor, and two or more of a lipid, an amino acid or amino acid derivative, an antioxidative agent, and an inorganic salt. Further provided are methods of using the growth medium, as well as kits and formulations of the growth medium. 1. A growth medium for culture of hematopoietic stem cells (HSCs) , comprising a basal medium and a supplement , the medium and/or supplement comprising a histone acetyltransferase (HAT) inhibitor , a histone deacetylase (HDAC) inhibitor and two or more of a lipid , an amino acid or amino acid derivative , an antioxidative agent , and an inorganic salt.2. The growth medium of claim 1 , wherein the basal medium is selected from OPTMIZER™ CTS™ T-Cell Expansion serum-free medium claim 1 , Dulbecco's Modified Eagle Media (DMEM) claim 1 , Iscove's Modified Dulbecco's Medium (IMDM) claim 1 , DMEM/F12 claim 1 , Advanced DMEM/F12 claim 1 , and KNOCKOUT™ DMEM/F12.3. The growth medium of claim 1 , wherein the HAT inhibitor is selected from 2 claim 1 ,6-Bis[(3-bromo-4-hydroxyphenyl)methylene]cyclohexanone claim 1 , MG149 claim 1 , C646 claim 1 , CPTH2 claim 1 , curcumin claim 1 , A-485 claim 1 , anacardic acid claim 1 , MB-3 claim 1 , and chalcones such as garcinol claim 1 , isogarcinol claim 1 , xanthohumol claim 1 , isoxanthohumol claim 1 , 2-hydroxycalchone claim 1 , 4-hydroxycalchone claim 1 , yakuchinone A claim 1 , and isoliquiritigenin.4. The growth medium of claim 1 , wherein the HAT inhibitor is present at a concentration of between 0.001 grams/liter (g/L) and 1 g/L.5. The growth medium of claim 4 , wherein the HAT inhibitor is present at a concentration of between 0.001 grams/liter (g/L) and 0.005 g/L.6. The growth medium of ...

EXPANSION AND DIFFERENTIATION OF STEM CELLS

The disclosure relates to the expansion and differentiation of mesenchymal stem cells and bone marrow cells, including retention of stem cell plasticity during expansion and differentiation of stem cells to produce osteocytes, chondrocytes and other cells of the mesodermal lineage. 1. A method for forming cells of mesodermal lineage from mesenchymal stem cells (MSC) comprising: (i) at least one differentiation factor for inducing formation of cells of mesodermal lineage from MSC; and', '(ii) tropoelastin,, 'contacting MSCs withwherein the number of cells of mesodermal lineage formed from MSC in the presence of tropoelastin is greater than the number of cells of mesodermal lineage formed in the absence of tropoelastin,thereby forming cells of mesodermal lineage from MSCs.2. The method of claim 1 , wherein the tropoelastin is arranged on a cell culture surface of a cell culture vessel to enable the MSCs to contact the tropoelastin when the MSCs are contacted with the cell culture surface.3. The method of claim 1 , wherein the tropoelastin is partially or fully solubilized in a cell culture medium for culture of an MSC.4. The method of claim 1 , wherein the method further comprises:(i) contacting MSCs with tropoelastin in the absence of factors that induce differentiation to induce proliferation of MSCs, thereby forming a population of MSCs; and(ii) contacting the population of MSCs with at least one differentiation factor for inducing formation of cells of mesodermal lineage from MSC and tropoelastin.5. The method of claim 1 , wherein the method further comprises:(i) culturing MSCs in a first medium containing tropoelastin to form a tropoelastin-cultured MSC population; and(ii) culturing said tropoelastin-cultured MSC population in a second medium, wherein the second medium includes at least one differentiation factor for inducing differentiation of an MSC.6. The method of claim 1 , wherein the tropoelastin is provided in the form of a complex with hyaluronic acid ...

METHODS AND COMPOSITIONS FOR MAKING ANTIBODIES AND ANTIBODY DERIVATIVES WITH REDUCED CORE FUCOSYLATION

The invention provides methods and compositions for preparing antibodies and antibody derivatives with reduced core fucosylation. 195-. (canceled)96. A population of humanized anti-CD70 antibodies , wherein the antibodies each comprise:{'sub': H', 'H', 'H, '(i) a humanized heavy chain comprising the three CDRs from SEQ ID NO:1 and a variable region framework sequence of human germline V1-2 or V1-18 and exon J-6, provided that any of positions H46, H67, H68, H69, H70, H71, H80, H81, H82, H82A and H91 (Kabat numbering) can be occupied by the amino acid occupying the corresponding position from SEQ ID NO:1,'}{'sub': κ', 'κ', 'κ, '(ii) a humanized light chain comprising the three CDRs from SEQ ID NO:2 and a variable region framework sequence of human germline Vexon B3 and Jexon J1, provided that any of positions L25 and L33 can be occupied by the amino acid occupying the corresponding position from SEQ ID NO:2, and'}(iii) an Fc domain, wherein at least 50% of the antibodies in the population of antibodies lack core fucosylation.97. The population of antibodies of claim 96 , wherein at least 70% of the antibodies in the population of antibodies lack core fucosylation.98. The population of antibodies of claim 96 , wherein position H46 of the antibodies is occupied by the amino acid occupying the corresponding position from SEQ ID NO:1.99. The population of antibodies of claim 96 , wherein at least one of positions H46 claim 96 , H67 claim 96 , H68 claim 96 , H69 claim 96 , H70 claim 96 , H71 claim 96 , H80 claim 96 , H81 claim 96 , H82 claim 96 , H82A and H91 (Kabat numbering) in the humanized heavy chain variable region of the antibodies is occupied by the residue occupying the corresponding position in SEQ ID NO:1.100. The population of antibodies of claim 96 , wherein at least one of positions L25 and L33 in the humanized light chain variable region of the antibodies is occupied by the residue occupying the corresponding position in SEQ ID NO:2.101. The population of ...

METHOD OF CULTURING IMMUNE CELLS, KIT FOR THEREOF, IMMUNE CELL CULTURED MEDIUM OBTAINED BY SAME METHOD, COSMETIC COMPOSITION AND PHARMACEUTICAL COMPOSITION COMPRISING THEREOF

The present invention relates to a technology for culturing Natural Killer cells (NK cells) applied to immunotherapy, and more particularly, a kit for adding to the serum-free immune cell culturing medium capable of effectively amplifying and activating immune cells that have been left for one day or longer after blood sampling or have become much weakened, while culturing NK cells to remarkably increase the portion of NK cells therein, compared with conventional immune cell-culturing methods, a method for culturing immune cells, a serum-free immune cell cultured medium obtained by the culturing method, and a cosmetic composition and a pharmaceutical composition comprising the cultured medium. 1. A kit for adding to a serum-free immune cell culturing medium comprisinga B unit composed of a basic solution comprising IL-2, L-glutamine and cell culture medium;a C1-1 unit composed of a cytokine 1-1 solution comprising IL-12 and IL-18 dissolved in the basic solution, wherein IL-12 is included at a concentration of 0.5-5 ng/mL and IL-18 is included at a concentration of 2-50 ng/mL;a C1-2 unit composed of a cytokine 1-2 solution comprising IL-12 and IL-18 dissolved in the basic solution, wherein IL-12 is included at a concentration of 5.1-15 ng/mL and IL-18 is included at a concentration of 30-120 ng/mL;a C2 unit composed of a cytokine 2 solution comprising IL-15 dissolved in the basic solution, wherein IL-15 is included at a concentration of 10-50 ng/mL;an A1 unit composed of an antibody 1 solution comprising an anti-CD16 and an anti-CD56 dissolved in the basic solution, wherein the anti-CD16 and the anti-CD56 are included at a concentration of 0.1-15 μg/mL each;an A2 unit composed of an antibody 2 solution comprising the antibody 1 solution and the basic solution, wherein a volume ratio of the antibody 1 solution:the basic solution is 1:6-10; anda D unit composed of an antibody-cytokine mixture solution comprising an anti-CD3 dissolved in the cytokine 1 solution, wherein ...

CELL CULTURE SUPPLEMENTS

The invention relates to cell culture supplements based on a platelet rich plasma fraction and a platelet poor plasma fraction. The supplements increase cell proliferation rate, improve selection of clonogenic cells, proliferation of cells from biopsies from elderly patients, maintenance of cell differentiation potential, and in vitro expansion of cell cultures also starting from an extremely low number of initially plated cells. The supplements may be freeze-dried and kept for long period of time as a quality controlled “off the shelf” product. 118.-. (canceled)19. A cell or tissue culture medium supplement consisting of:{'sup': '6', 'a) from 95% to 0.5% (volume/volume) of a platelet rich plasma fraction containing at least 2×10platelets/μL; and'}{'sup': '4', 'b) from 5% to 99.5% (volume/volume) of a platelet poor plasma fraction containing less than 5×10platelets/μL.'}20. The cell or tissue culture medium supplement according to claim 19 , wherein the cell or tissue culture medium supplement consists of:a) from 70% to 2.5% (volume/volume) of the platelet rich plasma fraction andb) from 30% to 97.5% (volume/volume) of the platelet poor plasma fraction.21. The cell or tissue culture medium supplement according to claim 19 , wherein the cell or tissue culture medium supplement consists of:75% (volume/volume) of the platelet rich plasma fraction and 25% (volume/volume) of the platelet poor plasma fraction, or50% (volume/volume) of the platelet rich plasma fraction and 50% (volume/volume) of the platelet poor plasma fraction, or20% (volume/volume) of the platelet rich plasma fraction and 80% (volume/volume) of the platelet poor plasma fraction, or10% (volume/volume) of the platelet rich plasma fraction and 90% (volume/volume) of the platelet poor plasma fraction.22. The cell or tissue culture medium supplement according to claim 19 , wherein the platelet rich plasma fraction contains from 5×10to 15×10platelets/μL.23. The cell or tissue culture medium supplement ...

CELL CULTURE MEDIA, KITS AND METHODS OF USE

Albumin-supplemented and xenogeneic product-free cell culture media, cell culture media supplements, and cell culture media kits for the support of primary culture of normal non-hematopoietic cells of mesodermal origin suitable for both research and clinical applications. 1. A cell culture medium for clinical growth of human adipose stromal cells for human clinical and therapeutic applications , said medium comprising:a basal medium suitable for mammalian cell culture;human albumin; andat least one of (i) growth promoting amounts of human insulin, (ii) human transferrin, (iii) human recombinant epidermal growth factor, (iv) human recombinant platelet-derived growth factor-BB.2. The cell culture medium of wherein the human source albumin is a recombinant protein derived from the human albumin gene expressed in an organism.3. The cell culture medium of claim 1 , wherein the insulin is a recombinant protein produced in a microorganism.4. The cell culture medium of claim 1 , wherein the human insulin (INS) is present in concentrations of between about 1 and 50 mg/l.5. The cell culture medium of claim 1 , wherein the transferrin is present in concentrations of between about 1 and 50 mg/l.6. The cell culture medium of claim 5 , wherein the transferrin is a recombinant protein produced in a microorganism.7. The cell culture medium of claim 1 , wherein the human source albumin is human plasma- or serum-derived albumin or recombinant human albumin in concentrations of between about 0.5 and 10 g/100 ml.8. The cell culture medium of claim 1 , wherein the recombinant human epidermal growth factor is in concentrations of between about 2 and 50 ng/ml.9. The cell culture medium of claim 1 , wherein the recombinant human platelet-derived growth factor-BB is in concentrations of between about 2 and 20 ng/ml.10. The cell culture medium of claim 1 , wherein the supplement further comprises at least one of endothelin-1 claim 1 , endothelin-2 and endothelin-3 claim 1 , in concentrations ...

CONDITIONED MEDIUM FROM HUMAN ADULT LIVER STEM CELLS AND ITS USE IN THE TREATMENT OF LIVER DISORDERS

The invention relates to cell-free compositions obtained by culturing adult-derived human liver stem/progenitor cells (ADHLSC) in cell culture medium and isolating the resulting conditioned medium (ADHLSC-CM) that has advantageous properties, such as anti-fibrotic effects. ADHLSC-CM, compositions based on ADHLSC-CM, and other related and derived products, can be used in cell culture processes or as a medicament, more particularly for the treatment of diseases involving organ injury, organ failure, in organ or cell transplantation or the pathological disruption, inflammation, degeneration, and/or proliferation of cells within a tissue or an organ, in particular within liver. 110-. (canceled)11. A method for producing a cell-free conditioned medium comprising the steps of culturing adult-derived human liver stem/progenitor cells (ADHLSC) in a cell culture medium and separating the cell culture medium from the cells.12. The method for producing a cell-free conditioned medium according to claim 11 , wherein:(a) the cell culture medium is a serum-free medium; and/or(b) the cell culture medium is separated from ADHLSC after culturing ADHLSC in the cell culture medium for at least 2 hours, at least 4 hours, at least 6 hours, at least 8 hours, at least 12 hours, or at least 24 hours; and/or(c) adult-derived human liver stem/progenitor cells (ADHLSC) co-express at least one mesenchymal marker selected from CD90, CD73, CD44, vimentin and a-smooth muscle actin with at least an hepatic marker selected from albumin, CD29, alpha-fetoprotein, alpha-1 antitrypsin, HNF-4 and MRP2 transporter.13. A method for producing a cell-free composition comprising Hepatocyte Growth Factor (HGF) claim 11 , Vascular Endothelial Growth Factor (VEGF) claim 11 , Eotaxin (CCL11) claim 11 , Interleukin-6 (IL-6) claim 11 , and Interleukin-8 (IL-8) at a concentration of at least 1 ng/ml comprising the steps of culturing adult-derived human liver stem/progenitor cells (ADHLSC) in a cell culture medium ...

METHODS FOR MODULATING MANNOSE CONTENT OF RECOMBINANT PROTEINS

The present invention relates to methods of modulating (e.g., reducing) the mannose content, particularly high-mannose content of recombinant glycoproteins. 123-. (canceled)24. A composition comprising an isolated antibody or an antigen-binding fragment thereof having low-mannose content.25. The composition according to claim 24 , wherein said isolated antibody or antigen-binding fragment having low-mannose content is an isolated human monoclonal antibody that binds IL-15 claim 24 , or an antigen-binding fragment thereof.26. The composition according to claim 25 , wherein said antibody comprises a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:4 claim 25 , or an amino acid sequence of at least 90% or at least 95% identity thereto claim 25 , or conservative amino acid substitutions thereof claim 25 , and a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:2 claim 25 , or an amino acid sequence of at least 90% or at least 95% identity thereto claim 25 , or conservative amino acid substitutions thereof.27. The composition according to claim 25 , wherein said antibody comprises a light chain variable region comprising one or more CDRs comprising amino acid sequences set forth in SEQ ID NOs:8-10 claim 25 , or an amino acid sequence of at least 90% or at least 95% identity thereto claim 25 , or conservative amino acid substitutions thereof claim 25 , and a heavy chain variable region comprising one or more CDRs comprising amino acid sequences set forth in SEQ ID NOs:5-7 claim 25 , or an amino acid sequence of at least 90% or at least 95% identity thereto claim 25 , or conservative amino acid substitutions thereof.28. The composition of claim 25 , further comprising a pharmaceutically acceptable carrier.29. A method of treating or preventing a disorder that is associated with overexpression of human IL-15 and/or in which a downregulation or inhibition of human IL-15 induced effects is beneficial ...

COMPOSITION FOR PROMOTING GROWTH OF STEM CELLS COMPRISING PHYTOSPHINGOSINE-1-PHOSPHATE OR DERIVATIVES THEREOF, AND COMPOSITION FOR CULTURING MEDIA OF STEM CELLS COMPRISING SAME

The present application relates to a composition for promoting proliferation of adult stem cells further comprising phytosphingosine-1-phosphate (P1P), O-cyclic P1P (cP1P), N-acetylphytosphingosine-1-phosphate (NAPS-1-P), and a pharmaceutically acceptable salt thereof in a basal medium, a composition for culturing adult stem cells comprising the same, or a composition for a serum-free or low-serum culture medium of adult stem cells. When the stem cells are cultured using the composition of the present application, the proliferation promotion, activity and differentiation capacity of the stem cells can be improved, and the death of the stem cells against external stress can be prevented. 14-. (canceled)5. A method selected from the group consisting of the following (i)-(iv):(i) culturing adult stem cells,(ii) promoting proliferation of adult stem cells,(iii) inhibiting cell death of adult stem cells, and(iv) promoting osteogenic differentiation of adult stem cellssaid method comprisingproviding adult stem cells; andculturing the adult stem cells in a medium containing P1P (phytosphingosine-1-phosphate), cP1P (O-cyclic P1P), and/or NAPS-1-P (N-acetyl phytosphingosine-1-phosphate), or a salt thereof.6. The method of claim 5 ,wherein the method is (i) culturing adult stem cells, andwherein the medium is animal blood serum free or contains 0.1 to 3% by weight of animal blood serum.7. The method of claim 5 ,wherein the method is (ii) promoting proliferation of adult stem cells, andwherein the medium is animal blood serum free or contains 0.1 to 3% by weight of animal blood serum.8. The method of claim 5 ,wherein the method is (iii) inhibiting cell death of adult stem cells, andwherein the medium is animal blood serum free or contains 0.1 to 3% by weight of animal blood serum.9. The method of claim 5 ,wherein the method is (iv) promoting osteogenic differentiation of adult stem cells, andwherein the medium is animal blood serum free or contains 0.1 to 3% by weight of ...

COMPOSITIONS AND METHODS FOR CULTURING AND EXPANDING CELLS

Provided herein are, inter alia, compositions, systems, kits, and methods for culturing and expanding cells (such as T cells, diploid or non-diploid cells), as well as methods for treating disorders (e.g., with T cells), and methods for producing biological molecules and compositions (e.g., proteins, viruses, viral particles or fragments thereof, etc.), including vaccines. 1. A serum-free cell culture medium composition or supplement composition comprising linoleic acid , at least one other omega-6 fatty acid , cholesterol , and a cyclodextrin , wherein the cholesterol is a synthetic cholesterol or an animal origin free cholesterol.2. The serum-free cell culture medium composition or supplement composition of claim 1 , wherein the at least one other omega-6 fatty acid is a polyunsaturated omega-6 fatty acid.38-. (canceled)9. The serum-free cell culture supplement composition of claim 2 , wherein the effective dilution of the supplement is from about 1:10 to about 1:5000.10. The serum-free cell culture medium composition or supplement composition of claim 2 , that is capable of culturing a cell that can produce a vaccine claim 2 , a virus claim 2 , a viral particle claim 2 , a viral protein or nucleic acid claim 2 , or a viral fragment.11. (canceled)12. The serum-free cell culture medium composition or supplement composition of claim 10 , wherein the cell is a bovine cell claim 10 , a canine cell claim 10 , a feline cell claim 10 , an insect cell claim 10 , an avian cell claim 10 , a primate cell or a human cell.1314-. (canceled)15. The serum-free cell culture medium composition or supplement composition of claim 10 , wherein the cell is selected from the group consisting of MRC-5 claim 10 , MRC-5 RCB claim 10 , MRC-9 claim 10 , WI-38 claim 10 , 2BS claim 10 , Walvax-2 claim 10 , IMR-90 claim 10 , IMR-91 claim 10 , KMB-17 claim 10 , HUT series cell claim 10 , Chang liver claim 10 , U937 claim 10 , MDCK claim 10 , CD4-expressing T cell claim 10 , CD8-expressing T cell ...

CELL CULTURE MEDIA COMPOSITIONS FOR PRIMARY CELLS

Platelet lysate compositions and cell culture media compositions for maintaining and/or growing mammalian cells, such as mammalian endothelial cells (ECs) and mammalian endothelial progenitor cells (EPCs), in particular human ECs (huECs) and human EPCs (huEPCs), such as primary huECs and primary huEPCs, are provided. The cell culture media compositions contain a basal medium, a platelet lysate and, optionally, one or more exogenously added growth factors. Also provided are methods for making and using such cell culture media compositions to grow and/or maintain ECs and EPCs, including huECs and huEPCs, as well as cell culture vessels, dishes, plates, and/or flasks pretreated with the cell culture media compositions.

METHODS AND COMPOSITIONS FOR MAKING ANTIBODIES AND ANTIBODY DERIVATIVES WITH REDUCED CORE FUCOSYLATION

The invention provides methods and compositions for preparing antibodies and antibody derivatives with reduced core fucosylation. 2. The culture medium of wherein Ris selected from the group consisting of —C≡CH claim 1 , —C≡CCH claim 1 , C(O)OCH claim 1 , —CHCN claim 1 , and —CHBr.3. The culture medium of wherein Ris —C≡CH and R-Ris —OAc.4. The culture medium of claim 1 , which is free of added animal protein.5. The culture medium of claim 1 , which is free of serum.6. The culture medium of claim 1 , which is free of added fucose.7. The culture medium of claim 1 , which is a powder.8. The culture medium of claim 1 , which is a liquid.9. The culture medium of claim 1 , wherein each of R-Ris independently selected from the group consisting of —OH and —OC(O)C-Calkyl.10. The culture medium of claim 1 , wherein each of R-Ris independently selected from the group consisting of —OH and —OAc.11. The culture medium of claim 1 , wherein Ris selected from the group consisting of —C≡CH and —C≡CCH.12. The culture medium of claim 1 , wherein Ris —C(O)OCH claim 1 , —CHCN claim 1 , or —CHBR.13. The culture medium of wherein the effective amount is an amount of the analog that is sufficient to decrease fucose incorporation into a complex N-glycoside-linked sugar chain of an antibody or antibody derivative by at least 90%.14. The culture medium of claim 1 , which:(i) is free of added animal protein;(ii) is free of serum;(iii) is free of added fucose; and(iv) is a powder or a liquid.15. The culture medium of claim 13 , which:(i) is free of added animal protein;(ii) is free of serum;(iii) is free of added fucose; and(iv) is a powder or a liquid.16. The culture medium of wherein the effective amount is an amount of the analog that is sufficient to decrease fucose incorporation into a complex N-glycoside-linked sugar chain of an antibody or antibody derivative by at least 90%.17. The culture medium of wherein the antibody or antibody derivative is a humanized or human antibody or ...

METHODS FOR MODULATING PRODUCTION PROFILES OF RECOMBINANT PROTEINS

The present invention relates to methods and compositions for modulating glycosylation of recombinant proteins expressed by mammalian host cells during the cell culture process. Also disclosed are methods of culturing a host cell expressing a recombinant protein in a cell culture medium comprising a disaccharide or a trisaccharide, while keeping the osmolality constant. 116-. (canceled)17. A method of producing a recombinant protein in fed-batch or batch mode , said method comprising culturing a mammalian host cell expressing said recombinant protein in a cell culture medium comprising a dissacharide or a trisaccharide , or supplemented with a dissacharide or a trisaccharide , while maintaining the osmolality similar to the one of a standard medium which does not comprise said disaccharide or trisaccharide.18. The method according to claim 17 , wherein said method increases the efficiency of at least one production run.19. The method according to claim 18 , wherein the efficiency of a production run is measured by an increase of the viable cell density and/or a lower decrease in cell viability.20. The method according to claim 17 , wherein the disaccharide is sucrose and the trisaccharide is raffinose.21. The method according to claim 17 , wherein the host cell is Chinese Hamster Ovary (CHO) cells.22. The method according to claim 17 , wherein the recombinant protein is selected from the group consisting of a recombinant fusion protein claim 17 , a growth factor claim 17 , a hormone claim 17 , a cytokine claim 17 , an antibody or antigen binding fragment thereof claim 17 , a human antibody or antigen-binding portion thereof claim 17 , a humanized antibody or antigen-binding portion thereof claim 17 , a chimeric antibody and antigen-binding portion thereof.23. The method according to claim 17 , wherein the concentration of disaccharide or trisaccharide in the cell culture medium is of about 0.1 mM to 100 mM.24. A method of culturing in fed-batch or batch mode a ...

CELL CULTURE MEDIUM AND CULTURE METHOD USING THE SAME

It is an object of the present invention to provide a cell culture medium capable of enhancing cell growth efficiency without using feeder cells, in particular which does not comprise serum. The present invention provides a cell culture medium which comprises growth arrest-specific 6 (GAS6) and does not comprise serum. 1. A cell culture medium , which comprises growth arrest-specific 6 (GAS6) , a basal medium , and at least one selected from the group consisting of decorin , matrix metalloproteinase-3 (MMP3) , osteopontin (OPN) , TNF-related weak inducer of apoptosis receptor (TWEAK R) , insulin-like growth factor-binding protein 2 (IGFBP2) , galectin 1 (LGALS1) , and insulin-like growth factor-1 (IGF-1) ,wherein the cell culture medium does not comprise serum, and the concentration of the GAS6 contained in the cell culture medium is between 2 ng/ml and 100 ng/ml.2. The cell culture medium according to claim 1 , wherein the cell culture medium supports the growth of iPS cells (induced pluripotent stem cells).3. The cell culture medium according to claim 1 , wherein the basal medium is BME medium claim 1 , BGJb medium claim 1 , CMRL1066 medium claim 1 , Glasgow MEM medium claim 1 , Improved MEM Zinc Option medium claim 1 , DMEM medium claim 1 , Medium 199 medium claim 1 , Eagle MEM medium claim 1 , aMEM medium claim 1 , DMEM medium claim 1 , Ham medium claim 1 , RPMI 1640 medium claim 1 , Fischer's medium claim 1 , or a mixed medium thereof.4. The cell culture medium according to claim 1 , wherein the basal medium is CHO-S-SFM Hybridoma-SFM claim 1 , eRDF Dry Powdered Media claim 1 , UltraCULTURE™ claim 1 , UltraDOMA™ claim 1 , UltraCHO™ claim 1 , and UltraMDCK™ claim 1 , Essential 8 claim 1 , STEMPRO hESC SFM claim 1 , mTeSR1 claim 1 , TeSR2 claim 1 , or ReproXF.5. The cell culture medium according to claim 1 , comprising decorin claim 1 , galectin 1 (LGALS1) claim 1 , and insulin-like growth factor-1 (IGF-1).6. The cell culture medium according to claim 1 , ...

MEDIA FOR CULTURING STEM CELLS

Well-defined, xeno-free culture media which comprise a TGF-beta isoform or the chimera formed between IL6 and the soluble IL6 receptor (IL6RIL6), which are capable of maintaining stem cells, and particularly, human embryonic stem cells, in an undifferentiated state are provided. Also provided are cell cultures comprising the culture media and the stem cells and methods of expanding and deriving embryonic stem cells in such well-defined, xeno-free culture media. In addition, the present invention provides methods of differentiating ESCs or EBs formed therefrom for the generation of lineage specific cells. 1. A culture medium comprising an IL6RIL6 chimera and basic fibroblast growth factor (bFGF) , wherein said IL6RIL6 chimera comprises amino acids 112-355 of GenBank Accession No. AAH89410 wherein said culture medium is capable of maintaining said human pluripotent stem cell in an undifferentiated state for at least 7 passages.2. The culture medium of claim 1 , wherein said IL6RIL6 chimera is provided at a concentration of at least 25 ng/ml.3. The culture medium of claim 1 , wherein said IL6RIL6 chimera is provided at a concentration of at least 200 ng/ml.4. The culture medium of claim 1 , wherein said culture medium is serum replacement-free.5. The culture medium of claim 1 , wherein said culture medium comprises serum replacement.6. The culture medium of claim 1 , wherein said culture medium is devoid of animal contaminant.7. A cell culture comprising a human pluripotent stem cell and a culture medium claim 1 , said culture medium comprising an IL6RIL6 chimera claim 1 , wherein said IL6RIL6 chimera comprises amino acids 112-355 of GenBank Accession No. AAH89410 claim 1 , wherein said culture medium is capable of maintaining said human pluripotent stem cell in an undifferentiated state.8. The cell culture of claim 7 , wherein said culture medium further comprises basic fibroblast growth factor (bFGF).9. The cell culture of claim 7 , wherein said IL6RIL6 chimera is ...

CELL CULTURE MEDIA

The present invention relates to dry cell culture media comprising amino acid components of certain particle size. Some dry powder cell culture media show poor dissolving properties and result in turbid solutions when they are dissolved in aqueous solutions. Using amino acid components of certain particle sizes significantly reduces that problem. 1. Dry powder cell culture medium in which more than 70% (w/w) of all cystein , cystin and tyrosine present in the medium is present in the form of particles with particle sizes below 50 μm and in which more than 70% (w/w) of all serine , isoleucine , leucine , glycine and phenylalanine present in the medium is present in the form of particles with particle sizes above 100 μm.2. Dry powder cell culture medium according to claim 1 , characterized in that more than 75% (w/w) of all arginine claim 1 , aspartic acid claim 1 , glutamic acid and threonine present in the medium is present in the form or particles with particle sizes of between 50 and 150 μm.3. Dry powder cell culture medium according to claim 1 , characterized in that the cell culture medium is a mammalian cell culture medium.4. Dry powder cell culture medium according to claim 1 , characterized in that the cell culture medium is a chemically defined cell culture medium.5. Dry powder cell culture medium according to claim 1 , characterized in that claim 1 , at least 70% of all cystein claim 1 , cystine and tyrosine present in the medium is present in the form of particles with particle sizes between 10 and 50 μm.6. Dry powder cell culture medium according to claim 1 , characterized in that at least 70% of all histidine present in the medium is present in the form of particles with particle sizes between 25 and 100 μm.7. Dry powder cell culture medium according to claim 1 , characterized in that 70% (w/w) of all other amino acids present in the medium beside cystein claim 1 , cystine claim 1 , tyrosine claim 1 , histidine claim 1 , arginine claim 1 , aspartic acid ...