Chondrogenic Progenitor Cells, Protocol For Derivation Of Cells And Uses Thereof

• US 20150240208A1
• Filed: 05/11/2015
• Published: 08/27/2015
• Est. Priority Date: 04/08/2010
• Status: Active Grant

• Current Assignee: University of Edinburgh
• Original Assignee: University of Edinburgh
• Inventors: Noble, Brendon Stewart, Pier, David Matthew
• Granted Patent: US 9,725,697 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 35/32 Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
• 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: A61P 19/04 for non-specific disorders of the connective tissue
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/84 from mammals
• CPC Class Codes: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2506/03 from non-embryonic pluripotent stem cells
• CPC Class Codes: C12N 5/0655 Chondrocytes; Cartilage
• CPC Class Codes: C12N 5/0662 Stem cells
• CPC Class Codes: G01N 33/5044 involving specific cell types

Abstract

The present invention provides an isolated population of chondrocyte precursor cells wherein 1% or less of the cells express Oct4, Nanog and/or TRA-1-60, 7% or less of the cells express no collagen II, collagen X, CD105 or Stro-1 and 85% or more of the cells express CBFA1, methods for preparing such cells and uses of chondrocyte cells derived from said precursor cells.

Claims

1. An isolated population of chondrocyte precursor cells wherein 1% or less of the cells express Oct4, Nanog and/or TRA-1-60, 7% or less of the cells express no collagen II, collagen X, CD105 or Stro-1 and 85% or more of the cells express CBFA1

6. An isolated population of chondrocyte precursor cells according to claim 1 or claim 2 for use in preparing chondrocyte cells for transplantation.

7. A method of treating a degenerative cartilage disease or cartilage injury comprising transplanting a population of chondrocyte precursor cells prepared from a population of isolated chondrocyte precursor cells according to claim 1 or claim 2 into a subject in need thereof.

8. A method of screening a compound for its ability to modulate chondrocyte growth, differentiation, or synthesis of cartilage components, by combining the compound with a population of chondrocyte precursor cells according to claim 1 or claim 2 and determining its effect.

9. A pharmaceutical composition for producing, repairing, or maintaining cartilage in vivo, containing a chondrocytes prepared from a chondrocyte precursor cell population according to claim 1 or claim 2.

2. An isolated population of chondrocyte precursor cells which are characterised by the loss of transdifferentiation potential.

5. A method for the preparation of a population of isolated chondrocytes comprising the steps of:

(a) resuspending a population of cells obtained by a method of claim 2 or claim 3 in a chondrogenic medium;

(b) culturing the cell suspension obtained in step (a).

3. A method for the preparation of chondrocyte precursor cells, comprising the steps of:

(a) differentiating a population of primate pluripotent stem (pPS) cells in chondrogenic media until the cells are greater than 75% confluent

(b) washing the cells and resuspending in a defined minimal growth medium

(c) culturing the cells obtained in (b) for a further period until the cells are differentiated into a population of chondrocyte precursor cells which are characterised by the loss of transdifferentiation potential.

4. A method for the preparation of chondrocyte precursor cells, comprising the steps of:

(a) differentiating a population of primate pluripotent stem (pPS) cells in chondrogenic media until the cells are greater than 75% confluent

(b) washing the cells and resuspending in a defined minimal growth medium

(c) culturing the cells obtained in (b) for a further period until the cells are differentiated into a population of chondrocyte precursor cells which are characterised by 1% or less of the cells express Oct4, Nanog and/or tra-1-60, 7% or less of the cells express no collagen II, collagen X, CD105 or Stro-1 and 85% or more of the cells express CBFA1

10. A method of administering a cellular composition comprising a chondrocyte lineage cell to a subject such that the cellular composition engrafts in the subject without generating an immune response to the cellular composition that would reject the engrafted cellular composition comprising 1) obtaining a cellular composition comprising chondrocyte lineage cell and 2) administering the chondrocyte lineage cell to the subject without administering an immuno-modulatory compound to the subject.