WO2008007082A2 - Milieu de croissance cellulaire - Google Patents

Milieu de croissance cellulaire Download PDF

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Publication number
WO2008007082A2
WO2008007082A2 PCT/GB2007/002584 GB2007002584W WO2008007082A2 WO 2008007082 A2 WO2008007082 A2 WO 2008007082A2 GB 2007002584 W GB2007002584 W GB 2007002584W WO 2008007082 A2 WO2008007082 A2 WO 2008007082A2
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WO
WIPO (PCT)
Prior art keywords
cell culture
embryonic stem
stem cells
cell
primate embryonic
Prior art date
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PCT/GB2007/002584
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English (en)
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WO2008007082A3 (fr
Inventor
Miho Furue
Peter Andrews
Tetsuji Okamoto
Denry J Sato
Original Assignee
University Of Sheffield
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University Of Sheffield filed Critical University Of Sheffield
Priority to AU2007274065A priority Critical patent/AU2007274065A1/en
Priority to JP2009518958A priority patent/JP5227318B2/ja
Priority to CA002657539A priority patent/CA2657539A1/fr
Priority to EP07766180A priority patent/EP2038404A2/fr
Publication of WO2008007082A2 publication Critical patent/WO2008007082A2/fr
Publication of WO2008007082A3 publication Critical patent/WO2008007082A3/fr
Priority to GB0823575A priority patent/GB2452456A/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0603—Embryonic cells ; Embryoid bodies
    • C12N5/0606—Pluripotent embryonic cells, e.g. embryonic stem cells [ES]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/05—Inorganic components
    • C12N2500/10—Metals; Metal chelators
    • C12N2500/20—Transition metals
    • C12N2500/24—Iron; Fe chelators; Transferrin
    • C12N2500/25—Insulin-transferrin; Insulin-transferrin-selenium
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/38—Vitamins
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/44—Thiols, e.g. mercaptoethanol
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/46—Amines, e.g. putrescine
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/90—Serum-free medium, which may still contain naturally-sourced components
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/10—Growth factors
    • C12N2501/115—Basic fibroblast growth factor (bFGF, FGF-2)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/90—Polysaccharides
    • C12N2501/91—Heparin

Definitions

  • the invention relates to the maintenance of primate embryonic stem cells, preferably human embryonic stem cells (hES), in culture in the absence of feeder cells and serum.
  • hES human embryonic stem cells
  • feeder cells typically fibroblasts which have been treated such that they cannot proliferate (e.g. mitomycin or irradiation treatment).
  • feeder fibroblasts are murine in origin but may be derived from other species
  • stem cell represents a generic group of undifferentiated cells that possess the capacity for self-renewal while retaining varying potentials to form differentiated cells and tissues.
  • Stem cells can be totipotent, pluripotent or multipotent. Derivative stem cells that have lost the ability to differentiate also occur and are termed 'nullipotenf stem cells.
  • a totipotent stem cell is a cell that has the ability to form all the cells and tissues that are found in an intact organism, including the extra-embryonic tissues (i.e. the placenta). Totipotent cells comprise the very early embryo (8 cells) and have the ability to form an intact organism.
  • a pluripotent stem cell is a cell that has the ability to form all tissues found in an intact organism although the pluripotent stem cell cannot form an intact organism.
  • a multipotent cell has a restricted ability to form differentiated cells and tissues.
  • adult stem cells are multipotent stem cells and are the precursor stem cells or lineage restricted stem cells that have the ability to form some cells or tissues and replenish senescing or damaged cells/tissues. Generally they cannot form all tissues found in an organism, although some reports have claimed a greater potential for such 'adult' stem cells than originally thought.
  • Pluripotent embryonic stem cells may be principally derived from two embryonic sources.
  • Cells isolated from the inner cell mass are termed embryonic stem (ES) cells.
  • ES embryonic stem
  • similar cells can be derived from the culture of primordial germ cells isolated from the mesenteries or genital ridges of days 8.5-12.5 post coitum embryos. These are referred to as embryonic germ cells (EG cells).
  • EG cells embryonic germ cells
  • Each of these types of pluripotential cell has a similar developmental potential with respect to differentiation into alternate cell types, but possible differences in behaviour (e.g. with respect to imprinting) have led these cells to be distinguished from one another.
  • the term "pluripotent embryonic stem cell” encompasses both cells derived from the inner cell mass and primordial germ cells.
  • WO96/22362 describes cell lines and growth conditions which allow the continuous proliferation of primate ES cells which exhibit a range of characteristics or markers which are associated with stem cells having pluripotent characteristics.
  • WO96/22362 discloses a method of maintaining primate ES cells in culture in an undifferentiated state in the presence of mouse fibroblast feeder cells and serum.
  • hES human ES
  • tissue engineering The potential utility of embryonic stem cells, particularly human ES (hES) cells, in therapeutic tissue engineering is well documented.
  • the pluripotent nature of these cells enables the selection and differentiation of hES cells into any cell/tissue type.
  • adventitious agents such as prions or viruses may infect the recipient when cells exposed to fetal bovine serum or murine feeder cells are used in therapy. It is therefore essential that cell culture of hES cells is conducted to minimise this risk.
  • feeder free and serum free conditions will help reduce this risk.
  • hES cells that have been differentiated into particular cell type derivatives have utility in the identification gene targets for new drugs and existing drugs since the cells are genotypically identical, stable and of known origin.
  • the use of ES cell lines of distinct genotypes also offers possible routes to drug screening and toxicology in a way pertinent to pharmacogenomics.
  • WOO 1/66697 discloses serum free growth of primate ES cells wherein the serum is replaced with fibroblast growth factor, typically human basic fibroblast growth factor (bFGF 4ng/ml).
  • fibroblast growth factor typically human basic fibroblast growth factor (bFGF 4ng/ml).
  • the cell culture media includes KnockOut SR ta (described in WO98/30679 which is incorporated by reference in its entirety) supplemented with bFGF.
  • the cell culture includes irradiated murine fibroblast feeder cells.
  • WO2006/029198 The development of a serum free and feeder free culture method for the growth of hES cells is disclosed in WO2006/029198. These growth conditions use elevated concentrations of bFGF (40-100ng/ml), supplemental agents that include gamma amino butyric acid, pipecholic acid and lithium and including amino acids, lipids, vitamins and glucose. WO2006/029198 also discloses the use of a cell culture substrate comprising human proteins such as fibronectin, vitronectin and laminin.
  • Furue et al discloses the serum and feeder free growth of mouse embryonic stem cells in the presence of leukaemia inhibitory factor (LIF). This is also described in WO2005/063968.
  • LIF leukaemia inhibitory factor
  • a method to maintain a primate embryonic stem cell in cell culture conditions that are cell feeder and serum free comprising: forming a preparation of primate embryonic stem cells in a cell culture vessel comprising cell culture medium that includes fibroblast growth factor, heparin and ascorbic acid, or ascorbic acid phosphate, or derivative thereof and maintaining the primate embryonic stem cells in an undifferentiated state.
  • a method to maintain primate embryonic stem cells in cell culture conditions that are cell feeder free and serum free comprising the steps: i) forming a preparation of primate embryonic stem cells in a cell culture vessel which is coated with a proteinaceous based cell culture support wherein said cells are cultured in a cell culture medium comprising: insulin, transferrin, sodium selenite, ethanolamine, 2-mercaptoethanol, oleic acid complexed with fatty acid free bovine albumin and further wherein the cell culture medium is supplemented with fibroblast growth factor, heparin and ascorbic acid, or ascorbic acid phosphate, or derivative thereof; and
  • pluripotent embryonic stem cells relates to both cells derived from the inner cell mass and primordial germ cells (EG). The possibility also exists of reprogramming somatic or extraembryonic differentiated cells, or more restricted stem cells back to a pluripotent state resembling that of ES cells derived from early embryos.
  • said cells have a stable karyotype.
  • ascorbic acid is ascorbic acid phosphate.
  • said primate embryonic stem cells are pluripotent human embryonic stem cells.
  • said primate embryonic stem cells retain the property to differentiate into at least the endoderm, mesoderm and ectoderm tissues throughout cell culture.
  • fibroblast growth factor is selected from the group consisting of: bFGF/FGF-2, hereinafter acidic FGF/FGF-1, bFGF, FGF-4, FGF-9, FGF-17 or FGF-18.
  • said fibroblast growth factor is bFGF.
  • bFGF is provided at a concentration of between l-50ng/ml; preferably about 10 ng/ml.
  • fibroblast growth factor is recombinant.
  • ascorbic acid phosphate is provided at a concentration of 10-300 ⁇ g/ml; preferably about lOO ⁇ g/ml.
  • 2-ethanolamine is provided at a concentration of 0.05-2. O ⁇ g/ml; preferably about 0.6 ⁇ g/ml.
  • oleic acid is provided at a concentration of 3-15 ⁇ g/ml; preferably about 9.5 ⁇ g/ml.
  • heparin is provided at a concentration of 10- 500ng/ml; preferably about lOOng/ml; preferably, heparin is heparin sulphate salt.
  • said proteinaceous cell culture support is collagen based.
  • the collagen-based cell culture support comprises type I collagen; preferably recombinant type I collagen.
  • said cell culture support comprises recombinant human proteins selected from the group consisting of: collagen I, collagen IV, fibronectin, laminin and vitronectin.
  • said cell support comprises at least two recombinant proteins selected from the group consisting of: collagen I, collagen IV, fibronectin, laminin and vitronectin.
  • said cell support comprises the recombinant proteins collagen I, collagen FV, fibronectin, laminin and vitronectin.
  • said cell culture support is Matrigel 1 " 1 .
  • said primate embryonic stem cells are passaged after addition of EDTA to the cell culture vessel.
  • said primate embryonic stem cells are passaged after addition of collagenase, preferably collagenase IV.
  • primate embryonic stem cells are passaged after addition of dispase.
  • primate embryonic stem cells are passaged after addition of trypsin/EDTA, preferably recombinant trypsin.
  • said primate embryonic stem cells are cloned.
  • the cell culture media does not include the buffering agent HEPES.
  • a method to differentiate primate embryonic stem cells into at least one cell-type in cell culture conditions that are cell feeder free and serum free comprising the steps: i) forming a preparation of primate embryonic stem cells in a cell culture vessel which is coated with a proteinaceous based cell culture support wherein said cells are cultured in a cell culture medium comprising: insulin, transferrin, sodium selenite, ethanolamine, 2-mercaptoethanol, oleic acid complexed with fatty acid free bovine albumin and further wherein the cell culture medium is supplemented with fibroblast growth factor and heparin; and ii) adding an agent that induces the differentiation of the primate embryonic stem cells into at least one cell-type.
  • the primate embryonic stem cells are human pluripotent embryonic stem cells.
  • the cell-type is a neurone.
  • the cell-type is an epithelial cell.
  • said proteinaceous based cell culture support is laminin.
  • a cell culture comprising: primate embryonic stem cells on a proteinaceous based cell culture support and cell culture media comprising: insulin, transferrin, sodium selenite, ethanolamine, 2- mercaptoethanol, oleic acid complexed with fatty acid free bovine albumin and further wherein the cell culture medium is supplemented with fibroblast growth factor, heparin and ascorbic acid, or ascorbic acid phosphate, or derivative thereof.
  • the primate embryonic stem cells are pluripotent human embryonic stem cells.
  • a cell culture comprising: primate embryonic stem cells on a proteinaceous based cell culture support and cell culture medium comprising: insulin, transferrin, sodium selenite, ethanolamine, 2- mercaptoethanol, oleic acid complexed with fatty acid free bovine albumin and further wherein the cell culture medium is supplemented with fibroblast growth factor and heparin characterised in that the cell culture further comprises at least one agent that induces differentiation of the primate embryonic stem cells into at least one cell-type.
  • the primate embryonic stem cells are pluripotent human embryonic stem cells.
  • a cell culture vessel comprising a cell culture medium that includes: insulin, transferrin, sodium selenite, ethanolamine, 2-mercaptoethanol, oleic acid complexed with fatty acid free bovine albumin and further wherein the cell culture medium is supplemented with fibroblast growth factor, heparin and ascorbic acid, or ascorbic acid phosphate, or derivative thereof.
  • said cell culture vessel further comprises primate embryonic stem cells; preferably pluripotent human embryonic stem cells
  • said vessel is selected from the group consisting of: a petri-dish; cell culture bottle or flask; multiwell plate.
  • "Vessel” is construed as any means suitable to contain a primate embryonic stem cell culture.
  • a cell culture medium container comprising a cell culture medium that includes: insulin, transferrin, sodium selenite, ethanolamine, 2-mercaptoethanol, oleic acid complexed with fatty acid free bovine albumin and further wherein the cell culture medium is supplemented with fibroblast growth factor, heparin and ascorbic acid, or ascorbic acid phosphate, or derivative thereof.
  • a cell culture medium container comprising a cell culture media that includes: insulin, transferrin, sodium selenite, ethanolamine, 2-mercaptoethanol, oleic acid complexed with fatty acid free bovine albumin and further wherein the cell culture medium is supplemented with fibroblast growth factor and heparin.
  • Figure 1 illustrates the effect of bFGF on human embryonic stem cell proliferation
  • Figure 2 illustrates the effect of bFGF and heparin on human embryonic stem cell proliferation and morphology
  • Figure 3 illustrates the expression of human embryonic stem cell markers in cells cultured in feeder free conditions
  • Figure 4 illustrates the growth of human embryonic stem cells in various medium
  • Figure 5 illustrates growth curves of human embryonic stem cell-line HUES in feeder free conditions
  • Figure 6 illustrates growth curves of human embryonic stem cell-line Shef 1 in feeder free conditions
  • Table 1 illustrates a summary of cell culture medium components for culturing human embryonic stem in feeder free conditions.
  • Hesf5 medium is identical to Hesf9 medium without the addition of oleic acid complexed with bovine albumin, ascorbic acid phosphate, bFGF, and heparin sulphate.
  • hESF9 medium ESF basal medium without HEPES supplemented with 9 factors, insulin, transferrin, sodium selenite, 2-mercaptoenthanol, 2-ethanolamine, oleic acid complexed with bovine albumin, ascorbic acid phosphate, bFGF, and heparin sulphate. 3. EDTA solution
  • Type I collagen (Nitta Gelatine, Co., Osaka, Japan)
  • the cells were seeded onto 25cm 2 flask coated with 100 ⁇ g/cm2 type I collagen in hESF9 medium.
  • hESF5 medium ESF basal medium without HEPES supplemented with 5 factors, insulin, transferrin, sodium selenite, 2-mercaptoenthanol and 2-ethanolamine.
  • Undifferentiated ES cells are harvested by EDTA solution. 3. Seed the cells onto laminin-coated dish in hESF5 supplemented with 10ng/ml bFGF and lOOng/ml heparin.
  • hESF5 medium supplemented with FA-BSA ESF basal medium without HEPES supplemented with 5 factors, insulin, transferrin, sodium selenite, 2- mercaptoethanol, 2-ethanolamine, and 0.5 mg/ml fatty acid free bovine albumin
  • Undifferentiated ES cells are harvested by EDTA solution.
  • Table 1 Defined medium for feeder and serum free growth (hESF9).

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Abstract

L'invention concerne un procédé pour cultiver des cellules souches embryonnaires de primate, dans les conditions sans chargeur ni sérum.
PCT/GB2007/002584 2006-07-12 2007-07-10 Milieu de croissance cellulaire WO2008007082A2 (fr)

Priority Applications (5)

Application Number Priority Date Filing Date Title
AU2007274065A AU2007274065A1 (en) 2006-07-12 2007-07-10 Cell growth medium
JP2009518958A JP5227318B2 (ja) 2006-07-12 2007-07-10 細胞増殖培地
CA002657539A CA2657539A1 (fr) 2006-07-12 2007-07-10 Milieu de croissance cellulaire
EP07766180A EP2038404A2 (fr) 2006-07-12 2007-07-10 Milieu de croissance cellulaire
GB0823575A GB2452456A (en) 2006-07-12 2008-12-29 Cell growth medium

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB0613756.6 2006-07-12
GBGB0613756.6A GB0613756D0 (en) 2006-07-12 2006-07-12 Cell culture medium

Publications (2)

Publication Number Publication Date
WO2008007082A2 true WO2008007082A2 (fr) 2008-01-17
WO2008007082A3 WO2008007082A3 (fr) 2008-03-06

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PCT/GB2007/002584 WO2008007082A2 (fr) 2006-07-12 2007-07-10 Milieu de croissance cellulaire

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EP (1) EP2038404A2 (fr)
JP (1) JP5227318B2 (fr)
CN (1) CN101490243A (fr)
AU (1) AU2007274065A1 (fr)
CA (1) CA2657539A1 (fr)
GB (2) GB0613756D0 (fr)
WO (1) WO2008007082A2 (fr)

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WO2010008815A1 (fr) 2008-06-24 2010-01-21 Bioactive Surgical, Inc. Sutures chirurgicales incoporées dans des cellules souches ou autres matériaux bioactifs
WO2011009613A1 (fr) * 2009-07-21 2011-01-27 Transgene Sa Composition enzymatique pour la digestion d'embryons de poulet
JP2013510567A (ja) * 2009-11-12 2013-03-28 テクニオン リサーチ アンド ディベロップメント ファウンデーション リミテッド 多能性幹細胞を未分化状態で培養する培地、細胞培養および方法
US8518431B2 (en) 2008-08-07 2013-08-27 Bioactive Surgical, Inc. Stem cell capture and immobilization coatings for medical devices and implants
EP2671945A1 (fr) * 2011-01-31 2013-12-11 National Institute of Biomedical Innovation Procédé de culture de cellules souches pluripotentes humaines
WO2014006379A1 (fr) * 2012-07-04 2014-01-09 University Of Edinburgh Culture de cellules souches comprenant des modulateurs de la voie de transduction du signal de protéine g
WO2015042356A1 (fr) * 2013-09-19 2015-03-26 The United States Of America, As Represented By The Secretary, Department Of Health & Human Services Milieu de culture chimiquement défini pour le maintien et la différenciation de cellules souches
US9040297B2 (en) 2006-08-02 2015-05-26 Technion Research & Development Foundation Limited Methods of expanding embryonic stem cells in a suspension culture
US20150329832A1 (en) * 2013-01-31 2015-11-19 Ajinomoto Co., Inc., Culture method for stable proliferation of pluripotent stem cell while maintaining undifferentiated state
KR20160012224A (ko) * 2013-05-30 2016-02-02 아지노모토 가부시키가이샤 줄기 세포의 배양용 배지
JP2016052305A (ja) * 2008-12-17 2016-04-14 ザ スクリプス リサーチ インスティテュート 幹細胞の作製と維持
EP3015551A4 (fr) * 2013-06-28 2016-11-23 Kaneka Corp Procédé de criblage de facteur de promotion de propagation de cellules souches pluripotentes
US9688956B2 (en) 2008-06-05 2017-06-27 National Cheng Kung University Method for preserving proliferation and differentiation potential of mesenchymal stem cells
US10385312B2 (en) 2005-08-29 2019-08-20 Technion Research & Development Foundation Limited Media for culturing stem cells
WO2019177420A1 (fr) * 2018-03-16 2019-09-19 사회복지법인 삼성생명공익재단 Composition de milieu comprenant du fgf -17 en tant qu'ingrédient efficace pour favoriser la prolifération de cellules souches et procédé de promotion de la prolifération de cellules souches à l'aide de celle-ci
WO2020004571A1 (fr) * 2018-06-27 2020-01-02 味の素株式会社 Additif pour culture de cellules souches, milieu de culture, et méthode de culture
US11697796B2 (en) 2017-01-23 2023-07-11 Stemcell Technologies Canada Inc. Media and methods for enhancing the survival and proliferation of stem cells
WO2023150293A3 (fr) * 2022-02-03 2023-09-14 Steakholder Foods Ltd. Formation accélérée de myotube

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CN111484970B (zh) * 2020-04-30 2022-09-16 广州再生医学与健康广东省实验室 一种低蛋白含量的无血清无饲养层胚胎与多能干细胞培养基

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998030679A1 (fr) * 1997-01-10 1998-07-16 Life Technologies, Inc. Substitut de serum pour cellules souches embryonnaires
WO2003029443A1 (fr) * 2001-09-28 2003-04-10 Es Cell International Pte Ltd Procedes de derivation et de propagation de cellules souches embryonnaires humaines (hes) non differenciees sur des matrices sans cellules nourricieres et sur des couches nourricieres humaines
WO2004111210A1 (fr) * 2003-06-12 2004-12-23 Yeda Research & Development Co.Ltd Amelioration de la differenciation d'oligodendrocytes
WO2006017370A2 (fr) * 2004-07-13 2006-02-16 Geron Corporation Milieu destine a la croissance de cellules souches embryonnaires humaines
WO2006029197A1 (fr) * 2004-09-08 2006-03-16 Wisconsin Alumni Research Foundation Milieu de culture et culture de cellules souches embryonnaires
WO2006036925A1 (fr) * 2004-09-28 2006-04-06 Wicell Research Institute, Inc. Culture de cellules souches embryonnaires de primates

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE466073T1 (de) * 2003-12-26 2010-05-15 Makoto Asashima Basalmedium zur kultivierung von es-zellen

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998030679A1 (fr) * 1997-01-10 1998-07-16 Life Technologies, Inc. Substitut de serum pour cellules souches embryonnaires
WO2003029443A1 (fr) * 2001-09-28 2003-04-10 Es Cell International Pte Ltd Procedes de derivation et de propagation de cellules souches embryonnaires humaines (hes) non differenciees sur des matrices sans cellules nourricieres et sur des couches nourricieres humaines
WO2004111210A1 (fr) * 2003-06-12 2004-12-23 Yeda Research & Development Co.Ltd Amelioration de la differenciation d'oligodendrocytes
WO2006017370A2 (fr) * 2004-07-13 2006-02-16 Geron Corporation Milieu destine a la croissance de cellules souches embryonnaires humaines
WO2006029197A1 (fr) * 2004-09-08 2006-03-16 Wisconsin Alumni Research Foundation Milieu de culture et culture de cellules souches embryonnaires
WO2006036925A1 (fr) * 2004-09-28 2006-04-06 Wicell Research Institute, Inc. Culture de cellules souches embryonnaires de primates

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
CALDWELL MAEVE A ET AL: "HEPARIN, BUT NOT OTHER PROTEOGLYCANS POTENTIATES THE MITOGENIC EFFECTS OF FGF-2 ON MESENCEPHALIC PRECURSOR CELLS" EXPERIMENTAL NEUROLOGY, ACADEMIC PRESS, NEW YORK, NY, US, vol. 152, no. 1, July 1998 (1998-07), pages 1-10, XP009073850 ISSN: 0014-4886 *
LI Y ET AL: "Expansion of human embryonic stem cells in defined serum-free medium devoid of animal-derived products" BIOTECHNOLOGY AND BIOENGINEERING, WILEY & SONS, HOBOKEN, NJ, US, vol. 91, no. 6, 20 September 2005 (2005-09-20), pages 688-698, XP002351170 ISSN: 0006-3592 *
REN WENWEN ET AL: "bFGF and heparin but not laminin are necessary factors in the mediums that affect NSCs differentiation into cholinergic neurons." NEUROLOGICAL RESEARCH JAN 2006, vol. 28, no. 1, January 2006 (2006-01), pages 87-90, XP002462651 ISSN: 0161-6412 *
ZHANG S-C ET AL: "In vitro differentiation of transplantable neural precursors from human embryonic stem cells" NATURE BIOTECHNOLOGY, NATURE PUBLISHING GROUP, NEW YORK, NY, US, vol. 19, no. 12, December 2001 (2001-12), pages 1129-1133, XP002268706 ISSN: 1087-0156 *

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US8876864B2 (en) 2008-06-24 2014-11-04 Bioactive Surgical, Inc Surgical sutures incorporated with stem cells or other bioactive materials
WO2010008815A1 (fr) 2008-06-24 2010-01-21 Bioactive Surgical, Inc. Sutures chirurgicales incoporées dans des cellules souches ou autres matériaux bioactifs
US8889168B2 (en) 2008-08-07 2014-11-18 Bioactive Surgical Inc. Stem cell capture and immobilization coatings for medical devices and implants
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JP2016052305A (ja) * 2008-12-17 2016-04-14 ザ スクリプス リサーチ インスティテュート 幹細胞の作製と維持
JP2017079802A (ja) * 2008-12-17 2017-05-18 ザ スクリプス リサーチ インスティテュート 幹細胞の作製と維持
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JP2017225456A (ja) * 2009-11-12 2017-12-28 テクニオン リサーチ アンド ディベロップメント ファウンデーション リミテッド 多能性幹細胞を未分化状態で培養する培地、細胞培養および方法
US20150240202A1 (en) * 2009-11-12 2015-08-27 Technion Research & Development Foundation Limited Culture media, cell cultures and methods of culturing pluripotent stem cells in an undifferentiated state
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US10745669B2 (en) 2013-01-31 2020-08-18 Ajinomoto Co., Ltd. Culture method for stable proliferation of pluripotent stem cell while maintaining undifferentiated state
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US11859205B2 (en) 2013-05-30 2024-01-02 Ajinomoto Co., Inc. Medium for culturing stem cells
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US11697796B2 (en) 2017-01-23 2023-07-11 Stemcell Technologies Canada Inc. Media and methods for enhancing the survival and proliferation of stem cells
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US20210214680A1 (en) * 2018-06-27 2021-07-15 Ajinomoto Co., Inc. Additive for culturing stem cells, culturing medium, and culturing method
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JP5227318B2 (ja) 2013-07-03
WO2008007082A3 (fr) 2008-03-06
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GB2452456A (en) 2009-03-04
GB0613756D0 (en) 2006-08-23

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