Objective:To explore the possibility of the transfection of MyoD gene induced bone marrow mesenchymal stem cells(MSCs) to differentiate into myoblasts in vitro.Methods:The eukaryotic expression plasmid vector pIRES2-EGRP-MyoD was transfected into MSCs with lipotransfection method,and the positive cells were selected by G418;The expression of MyoD was detected in the transfected MSCs with RT-PCR and the amplified,purified product was identified by sequencing;The reporter gene enhanced green fluorescence protein(EGFP)was observed in the transfected cells under a fluorescent and a laser confocal microscopes;Immunohistochemical methods was used to examine the expressions of MyoD,myogenin,myosin,myoglobin and desmin in the differentiated,cells.The ultrastructure changes of the cells before and after transfection were observed with electron microscopy.Results:The expression of MyoD was detected in the transfected MSCs with RT-PCR and the amplified,purified product was as same in sequence as thwat from Genbank;Green fluorescence was observed in the transfected cells under a fluorescent and a laser confocal microscopes;Immunohistochemical methods indicated that MyoD,myogenin,myosin,myoglobin and desmin were expressed in the transfected cells;The transfected cells showed the morphologcal characteristics of matrue cells with filaments in their cytoplasm.Conclusion:MyoD gene can induce cultured MSCs to successfully differentiate into myoblasts,probably providing an experimental foundation for trauma repair.
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