Study On Chondrogenesis Of Bone Marrow Mesenchymal Stem Cells Under Induction Of Cartilage-derived Morphogenetic Protein-2 In Vitro

ObjectiveTo study the cartilage differentiation of mouse bone marrow mesenchymal stem cells(BMSCs) induced by cartilage-derived morphogenetic protein-2 in vitro.MethodsThe BMSCs were isolated from mouse bone marrow and cultured in vitro.The cells in passage 3 were chosen to induce into chondrogenic differerntiation with different culture media.In the control group(group A),the cells were cultured in L-DMEM containing 10%fetal bovine serum.In the experimental group,the cells were cultured in L-DMEM containing 10%fetal bovine serum with CDMP-2 100ng/ml(group B),TGF-β1 10ng/ml(group C) and CDMP-2 100ng/ml+TGF-β1 10ng/ml(group D).Under induction for 14 days,morphology of cells was observed under phase-contrast microscopy.Type-Ⅱcollagen protein was examined by using immunohistochemistry staining and western blot.The sulfate glycosaminoglycan was measured by alkaline phosphatase toluidine blue staining.ResultsUnder the induction of culture media in groups B,C,D,the morphology of cells changed from spindle appearance to the typical chondrocyte-like polygon shape.GAGs was distributed evenly in the cytoplasm and type-Ⅱcollagen protein was positive in groups B,C,D and negative in group A.Type-Ⅱcollagen protein in CDMP-2 100ng/ml +TGF-β1 10ng/ml(group D) was more than either CDMP-2 100ng/ml(group B) or TGF-β1 10ng/ml(group C).There was statistically significant difference in groups.ConclusionMouse bone marrow mesenchymal stem cells can be differentiated into chondrogenic phonotype with the induction of CDMP-2 in vitro.By comparison, CDMP-2 was more effectively than TGF-β1 and the combination of CDMP-2 and TGF-β1 was more effectively than either CDMP-2 or TGF-β1 alone.