Coª²expression of both green fluorescent protein and myogenic regulatory factors in bone marrowª²derived mesenchymal stem cells

    LI Meiª²Shan, ZHANG Cheng, CHEN Songª²Lin, YU Meiª²Juan, ZHANG Yaª²Ni, WANG Shuª²Hui, XIONG Fu

    Department of Neurology, First Affiliated Hospital, Research Center for Stem Cell and Tissue Engineering, Sun Yatª²sen University, Guangzhou 510080, China

    ¡¾Abstract¡¿ AIM: To investigate the transfection of bone marrowª²derived mesenchymal stem cells (MSCs) by recombinant adenoª²virus (Ad), and the expressions of both green fluorescent protein (GFP) and myogenic regulatory factors, which are MyoD and Myogenin. METHODS: Rat MSCs were separated, cultured and expanded in vitro, and taken for identification of surface antigens by flow cytometry (FCM). Adª²GFP was amplified and identified to transfect MSCs. Expressions of both MyoD and Myogenin were detected in transfected MSCs by RTª²PCR. Differentiation of MSCs was induced by cocultured with C2C12 myoblasts, and MyoD expression was identified by immunofluorescence (IF). RESULTS: FCM showed that CD11b and CD45 were negative, CD29 and CD44 were positive in MSCs surface antigens. With the increasing of Adª²GFP multiple of infection (MOI), transfection rate were also increased. When MOI was over 100, cytopathic effect appears on MSCs. Expressions of both MyoD and Myogenin in transfected MSCs were approved by RTª²PCR; and MyoD protein can be found in induced MSCs by IF. CONCLUSION: MSCs can be effectively transfected by Adª²GFP and express GFP; and activation of intrinsic myogenic regulatory factors will enhance the myogenic differentiation of MSCs. ......