Chondrogenic Differentiation of Mouse Bone Marrow Mesenchymal Stem Cells Induced by Cartilage-derived Morphogenetic Protein-2 In Vitro

To study the cartilage differentiation of mouse mesenchymal stem cells(MSCs) induced by cartilage-derived morphogenetic proteins-2 in vitro,the MSCs were isolated from mouse bone marrow and cultured in vitro.The cells in passage 3 were induced into chondrogenic differentiation with different concentrations of recombinant human cartilage-derived morphogenetic proteins-2(0,10,20,50 and 100 ng/mL).After 14 days of induction,morphology of cells was observed under phase-contrast microscope.Collagen Ⅱ mRNA and protein were examined with RT-PCR,Western blotting and immunocytochemistry respectively and the sulfate glycosaminoglycan was measured by Alcian blue staining.RT-PCR showed that CDMP-2 could promote expression of collagen Ⅱ mRNA in an dose-dependant manner,especially at the concentration of 50 ng/mL and 100 ng/mL.Immunocytochemistry and Western blotting revealed a similar change.Alcian blue staining exhibited deposition of typical cartilage extracellular matrix.Our results suggest that mouse bone marrow mesencymal stem cells can differentiate into chondrogenic phonotype with the induction of CDMP-2 in vitro,which provides a basis for further research on the role of CDMP-2 in chondrogenesis.