/ METHODS AND COMPOSITIONS FOR GENERATING CHONDROCYTE LINEAGE CELLS AND/OR CARTILAGE LIKE TISSUE

摘要

a. A primitive line-like mesoderm population, optionally, CD56+, PDGFRalpha+ KDR- primitive line-like mesoderm population i. FGF agonists; ii. BMP inhibitors; optionally noggin, LDN-193189, dosomorphine; and iii. optionally, a TGFbeta inhibitor, optionally SB431524; and a Wnt inhibitor, optionally comprising one or more of DKK1, IWP2, or XAV939, to characterize a periaxial mesoderm population expressing cell surface CD73, CD105 and/or PDGFR-beta by incubating with a periaxial mesoderm specification cocktail. ; b. i. culturing the periaxial mesoderm population expressing CD73, CD105 and/or PDGFR-beta at high cell density, optionally in serum-free or serum-containing medium; ii. culturing a high cell density CD73+, CD105+ and/or PDGFRbeta+ periaxial mesoderm population in serum-free medium with a TGFbeta3 agonist to produce a high cell density Sox9+, collagen 2+ chondrocyte progenitor population , generating a chondrocyte progenitor population; and c. i. culturing a high cell density Sox9+, collagen 2+ chondrocyte precursor population with a TGFbeta3 agonist for a long period of time to produce joint-like non-hypertrophic chondrocytes and/or cartilage-like tissue; or ii. Chondrocytes and/or cartilage, optionally comprising the step of culturing a high cell density Sox9+, Collagen2+ chondrocyte precursor population with a BMP4 agonist for an extended period of time to produce hypertrophic chondrocyte-like cells and/or cartilage-like tissue A method of generating joint-like non-hypertrophic chondrocytes and/or cartilage-like tissue and/or hypertrophic chondrocyte-like cells and/or cartilage-like tissue.