Chondrocyte precursors derived from human embryonic stem cells

• US 20060148077A1
• Filed: 02/01/2006
• Published: 07/06/2006
• Est. Priority Date: 12/07/2001
• Status: Active Grant

• Current Assignee: Asterias Biotherapeutics Incorporated (Lineage Cell Therapeutics, Inc.)
• Original Assignee: Thies R. Scott
• Inventors: Thies, R. Scott
• Granted Patent: US 7,906,330 B2
• Time in Patent Office: Days
• US Class Current: 435/363
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: C12N 2500/44 Thiols, e.g. mercaptoethanol
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/395 Thyroid hormones
• CPC Class Codes: C12N 2501/41 Hedgehog proteins; Cyclopamine (inhibitor)
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0655 Chondrocytes; Cartilage

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

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. The cell population of claim 1, in which less than 1% of the cells synthesize elastin, Type I collagen, Type X collagen, or osteocalcin.

3. The cell population of claim 1, which causes closure of a 6 mm hole within 2 months in the animal model described by Koppel et al., Biomaterials 22:

1407, 2001.

4. 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 the cells of claim 1.

5. The cell population of claim 1, comprising less than 1% undifferentiated pPS cells.

6. The cell population of claim 1, comprising cells that have been genetically altered to express telomerase reverse transcriptase (TERT) at an elevated level.

7. Two cell populations, comprising the differentiated cell population of claim 1, and the undifferentiated pPS cell line from which it was obtained.

8. A method for obtaining the cell population of claim 1, comprising differentiating the pPS cells, then culturing the differentiated cells with a mixture of chondrocyte differentiation factors.

9. The method of claim 8, comprising differentiating pPS cells to form embryoid bodies.

10. The method of claim 8, 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).

11. A method for producing cartilage, comprising incubating the cell population of claim 1 under conditions where the cell population synthesizes either Type II collagen or aggrecan.

12. A pharmaceutical composition for producing, repairing, or maintaining cartilage in vivo, comprising the cell population of claim 1 in a physiologically compatible excipient.

13. The pharmaceutical composition of claim 12, further comprising a matrix that is reabsorbed in vivo.

14. 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 cell population of claim 1, 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.

15. A method of reconstructing cartilage in a subject, comprising administering to the subject the cell population of claim 1.