KELLER, Gordon,CRAFT, April M. M,．克拉夫特,戈登.凱勒,阿普里爾.,.克拉夫特
申请号：
HK16107222.7
公开号：
HK1219293A
申请日:
2016.06.22
申请国别(地区):
HK
年份:
2017
代理人:
摘要：
A method for generating chondrocytes and/or cartilage, optionally articular like non-hypertrophic chondrocyte cells and/or cartilage like tissue and/or hypertrophic chondrocyte like cells and/or cartilage like tissue, the method comprising: a. culturing a primitive streak-like mesoderm population, optionally a CD56+, PDGFRalpha+ KDR-primitive streak-like mesoderm population, with a paraxial mesoderm specifying cocktail comprising: i. a FGF agonist； ii. a BMP inhibitor； optionally Noggin, LDN-193189, Dorsomorphin； and iii. optionally one or more of a TGFbeta inhibitor, optionally SB431524； and a Wnt inhibitor, optionally DKK1, IWP2, or XAV939； to specify a paraxial mesoderm population expressing cell surface CD73, CD105 and/or PDGFR-beta； b. generating a chondrocyte precursor population comprising: i. culturing the paraxial mesoderm population expressing CD73, CD105 and/or PDGFR-beta at a high cell density optionally in serum free or serum containing media； ii. culturing the high cell density CD73+, CD105+ and/or PDGFRbeta+ paraxial mesoderm population with a TGFbeta3 agonist in serum free media to produce a high cell density Sox9+, collagen 2+ chondrocyte precursor population； and c. either i. culturing the high cell density Sox9+, collagen 2+ chondrocyte precursor population with the TGFbeta3 agonist for an extended period of time to produce an articular like non-hypertrophic chondrocyte cells and/or cartilage like tissue； or ii. culturing the high cell density Sox9+ collagen2+ chondrocyte precursor population with a BMP4 agonist for an extended period of time to produce a hypertrophic chondrocyte like cells and/or cartilage like tissue.
