WO2003050250B1 - Chondrocyte precursors derived from human embryonic stem cells - Google Patents

Chondrocyte precursors derived from human embryonic stem cells

Info

Publication number
WO2003050250B1
WO2003050250B1 PCT/US2002/039090 US0239090W WO03050250B1 WO 2003050250 B1 WO2003050250 B1 WO 2003050250B1 US 0239090 W US0239090 W US 0239090W WO 03050250 B1 WO03050250 B1 WO 03050250B1
Authority
WO
WIPO (PCT)
Prior art keywords
cells
cell population
differentiated cell
population according
differentiated
Prior art date
Application number
PCT/US2002/039090
Other languages
French (fr)
Other versions
WO2003050250A3 (en
WO2003050250A2 (en
Inventor
R Scott Thies
Original Assignee
Geron Corp
R Scott Thies
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 Geron Corp, R Scott Thies filed Critical Geron Corp
Priority to IL16213202A priority Critical patent/IL162132A0/en
Priority to EP02804739A priority patent/EP1463800A4/en
Priority to CA2468335A priority patent/CA2468335C/en
Priority to GB0414956A priority patent/GB2399348A/en
Priority to AU2002366602A priority patent/AU2002366602B2/en
Priority to JP2003551272A priority patent/JP2005511083A/en
Publication of WO2003050250A2 publication Critical patent/WO2003050250A2/en
Publication of WO2003050250A3 publication Critical patent/WO2003050250A3/en
Publication of WO2003050250B1 publication Critical patent/WO2003050250B1/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
    • C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
    • C12N5/0655—Chondrocytes; Cartilage
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • 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
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • 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
    • 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
    • C12N2501/10—Growth factors
    • C12N2501/15—Transforming growth factor beta (TGF-β)
    • 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
    • C12N2501/30—Hormones
    • C12N2501/38—Hormones with nuclear receptors
    • C12N2501/395—Thyroid hormones
    • 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
    • C12N2501/40—Regulators of development
    • C12N2501/41—Hedgehog proteins; Cyclopamine (inhibitor)
    • 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
    • C12N2506/00—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells
    • C12N2506/02—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from embryonic cells
    • 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
    • C12N2510/00—Genetically modified cells

Abstract

This invention provides a system for obtaining cells of the chondrocyte lineage by differentiating primate pluripotent stem cells. The process involves culturing the cells as a micromass or other aggregate form in a cocktail of differentiation agents that facilitates outgrowth of the desired cell type. Progeny are capable of synthesizing Type II collagen or aggrecan, or other products that are characteristic of the chondrocyte lineage. Chondrocytes and chondrocyte precursor cells obtained according to this disclosure are suitable for use in both research and clinical therapy.

Claims

AMENDED CLAIMS[Received by the International Bureau on 12 August 2003 (12.08.03): original claims 1-20 replaced by amended claims 1-21]
1. A cell population obtained by differentiating primate pluripotent stem (pPS) cells, in which at least 5% of the cells synthesize either Type II collagen or aggrecan from an endogenous gene.
2. A cell population obtained by differentiating a line of primate pluripotent stem (pPS) cells as part of a system for generating chondrocyte lineage cells, wherein at least 5% of the differentiated cells synthesize either Type II collagen or aggrecan from an endogenous gene, and wherein the system further comprises the line of undifferentiated pPS cells from which the differentiated cells were produced.
3. A system for generating pancreatic hormone secreting cells, comprising the differentiated cell population of claim 1 , and the undifferentiated pPS cell line from which it was obtained.
4. , The differentiated cell population according to any preceding claim, in which less than 1% of the cells synthesize elastin, Type I collagen, Type X collagen, or osteocaicin.
5. The differentiated cell population according to any of claims 1-3, which causes closure of a 6 mm hole within 2 months in the animal model described by Koppel et al., Biomaterials 22:1407, 2001.
6. A population of chondrocyte progenitors that proliferates in an in vitro culture, obtained by differentiating primate pluripotent stem (pPS) cells, and capable of forming progeny having the characteristics of differentiated cells according to claims 1 -3.
7. The differentiated cell population according to any of claims 1-3, comprising less than 1% undifferentiated pPS cells.
8. The differentiated cell population according to any of claims 1-3, comprising cells genetically altered to express telomerase reverse transcriptase (TERT) at an elevated level.
9. The differentiated cell population according to any of claims 1 -3, which has the same genome as an established line of primate embryonic stem cells.
22
10. A method for obtaining the differentiated cell population according to any of claims 1-3, comprising differentiating the pPS cells, then culturing the differentiated cells with a mixture of chondrocyte differentiation factors.
11. The method of claim 9, comprising differentiating pPS cells to form embryoid bodies.
12. The method of claim 9, wherein the mixture of chondrocyte differentiation factors comprises one or more factors selected from transforming growth factors (TGF), fibroblast growth factors (FGF), growth and differentiation factors (GDF), bone morphogenic proteins (BMP), hedgehog proteins (HH), L-ascorbic acid, and parathyroid hormone-related protein (PTHrP).
13. A method for producing cartilage, comprising incubating the differentiated cell population according to any of claims 1 -3 under conditions where the cell population synthesizes either Type II collagen or aggrecan.
14. A pharmaceutical composition for producing, repairing, or maintaining cartilage in vivo, comprising the differentiated cell population according to any of claims 1-3 in a physiologically compatible excipient.
15. The pharmaceutical composition of claim 13, further comprising a matrix that is reabsorbed in vivo.
16. A method of screening a compound for its ability to modulate chondrocyte growth, differentiation, or synthesis of cartilage components, comprising combining the compound with the differentiated cell population according to any of claims 1-3, determining any phenotypic or metabolic changes in the cell population that result from being combined with the compound, and correlating the change with an ability of the compound to modulate chondrocyte growth, differentiation, or synthesis of cartilage components.
17. A method of reconstructing cartilage in a subject, comprising administering to the subject a differentiated cell population according to claims 1-3.
18. Use of the differentiated cell population according to any of claims 1-3 in the preparation of a medicament for reconstructing cartilage in a subject.
19. Use of the differentiated cell population according to any of claims 1 -3 in the preparation of a medicament for the treatment of joint trauma, arthritis, or osteoarthritis, or for use in cosmetic surgery.
20. The differentiated cell population according to any of claims 1 -3, wherein the pPS cells are progeny of cells obtained from a human blastocyst.
21. The differentiated cell population according to any of claims 1 -3, wherein the pPS cells are human embryonic stem cells.
24
PCT/US2002/039090 2001-12-07 2002-12-06 Chondrocyte precursors derived from human embryonic stem cells WO2003050250A2 (en)

Priority Applications (6)

Application Number Priority Date Filing Date Title
IL16213202A IL162132A0 (en) 2001-12-07 2002-12-06 Chondrocyte precursors derived from human embryonic stem cells
EP02804739A EP1463800A4 (en) 2001-12-07 2002-12-06 Chondrocyte precursors derived from human embryonic stem cells
CA2468335A CA2468335C (en) 2001-12-07 2002-12-06 Chondrocyte precursors derived from human embryonic stem cells
GB0414956A GB2399348A (en) 2001-12-07 2002-12-06 Chondrocyte precursors derived from human embryonic stem cells
AU2002366602A AU2002366602B2 (en) 2001-12-07 2002-12-06 Chondrocyte precursors derived from human embryonic stem cells
JP2003551272A JP2005511083A (en) 2001-12-07 2002-12-06 Chondrocyte progenitor cells derived from human embryonic stem cells

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US33904301P 2001-12-07 2001-12-07
US60/339,043 2001-12-07

Publications (3)

Publication Number Publication Date
WO2003050250A2 WO2003050250A2 (en) 2003-06-19
WO2003050250A3 WO2003050250A3 (en) 2004-02-12
WO2003050250B1 true WO2003050250B1 (en) 2004-04-15

Family

ID=23327241

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2002/039090 WO2003050250A2 (en) 2001-12-07 2002-12-06 Chondrocyte precursors derived from human embryonic stem cells

Country Status (10)

Country Link
US (5) US20030109038A1 (en)
EP (1) EP1463800A4 (en)
JP (1) JP2005511083A (en)
KR (1) KR100973453B1 (en)
CN (1) CN1599793A (en)
AU (1) AU2002366602B2 (en)
CA (1) CA2468335C (en)
GB (1) GB2399348A (en)
IL (1) IL162132A0 (en)
WO (1) WO2003050250A2 (en)

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Publication number Publication date
US7906330B2 (en) 2011-03-15
AU2002366602B2 (en) 2008-07-03
JP2005511083A (en) 2005-04-28
KR20050044733A (en) 2005-05-12
US20190024053A1 (en) 2019-01-24
WO2003050250A3 (en) 2004-02-12
KR100973453B1 (en) 2010-08-02
US20030109038A1 (en) 2003-06-12
GB2399348A (en) 2004-09-15
US20140134721A1 (en) 2014-05-15
CA2468335A1 (en) 2003-06-19
US8546101B2 (en) 2013-10-01
US20060148077A1 (en) 2006-07-06
CA2468335C (en) 2014-07-08
IL162132A0 (en) 2005-11-20
WO2003050250A2 (en) 2003-06-19
GB0414956D0 (en) 2004-08-04
US20110129867A1 (en) 2011-06-02
EP1463800A4 (en) 2006-04-26
AU2002366602A1 (en) 2003-06-23
EP1463800A2 (en) 2004-10-06
CN1599793A (en) 2005-03-23

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