Study On Chondrogenesis Of Myoblast Under Transfection Of Cartilage-Derived Morphogenetic Protein-2in Vitro

Objective1. To study the feasibility cartilage differentiation of mouse myoblastcells transferred by cartilage-derived morphogenetic protein-2in vitro.2. To study the feasibility of which CDMP-2transfer mouse myoblastcells,and the specificity expression after the transfection.MethodsThe mouse myoblast were cultured in vitro to passage.To carry out thecells in passage3to experimental treatment. In the control group (groupA),the mouse myoblasts were cultured in L-DMEM without fetal bovineserum;In the experimental group B,the mouse myoblasts with sodiumhyaluronate were cultured in L-DMEM without fetal bovine serum;In theexperimental group C,the mouse myoblasts with CDMP-2100ng/ml werecultured in L-DMEM without fetal bovine serum; In the experimental groupD,the mouse myoblast were transfected by CDMP-2were cultured inL-DMEM without fetal bovine serum.Under culture after16days, morphologyof cells were observed under phase-contrast microscopy.Theglycosaminoglycan was measured by toluidine blue staining.Type-Ⅱcollagenprotein was examined by using immunohistochemistry staining and westenblot. ResultsUnder the culture after16days,the morphology of cells in induce groupand transfer group changed from spindle apperaance to the typicalchondrocyte-like polygon shape.GAGs was distributed evenly in thecytoplasm and type-Ⅱcollagen protein has stainde by SP was positive ingroups C,D and negative in group A,B. There was statistically significantdifference in groups.ConclusionMouse myoblast cells can be differentiated into chondrogenic phonotypewith the induction of CDMP-2in vitro.The transferred mouse myoblast cellscan excrete the specificity protein and polysaccharide of cartilage cells.Bycomparison, transfection was more effectively than induction.