Primary murine limb bud mesenchymal cells in long-term culture complete chondrocyte differentiation: TGF-β delays hypertrophy and PGE2 inhibits terminal differentiation

摘要: In vitro models of endochondral bone formation using both primary and immortalized cells have provided insight regarding factors signaling pathways involved in chondrocyte maturation formation. However, murine cell culture differentiation not been established but enormous potential due to the possible use from transgenic knockout animals. Here, we show that stage E11.5 embryonic limb bud mesenchymal stem micromass progress through stages chondrogenesis, hypertrophy, terminal differentiation, matrix calcification. This system recapitulated sequential expression genes characterize including Sox9, col2, colX, MMP13, VEGF, osteocalcin. TGF-beta treatment for up 21 days markedly delayed rate inhibited calcification its related gene expression. TGF-beta-treated cultures, hypertrophic markers osteocalcin were reduced or absent. PGE2 had minimal effects on hypertrophy Thus, sequentially differentiate various establish a model whereby role specific molecules can be examined derived mice.