直接与间接接触共培养对诱导骨髓间充质干细胞向心肌样细胞分化的影响

Effect of the differentiation of bone mesenchymal stem cells into cardiomyocyte-like cells induced by direct and indirect contact co-culture

目的研究不同接触方法共培养对骨髓间充质干细胞(BMSCs)呈心肌样细胞分化的影响。方法通过体外扩增BMSCs,在细胞水平上研究心肌细胞直接接触共培养、transwell小室建立的间接接触共培养模拟的心肌微环境,对BMSCs向心肌细胞分化的影响。通过倒置显微镜观察形态,2 w后运用免疫荧光、RT-PCR等技术,检测两试验组细胞心肌特异性蛋白:心肌肌钙蛋白T(c Tn T)表达情况。结果免疫荧光及RT-PCR结果均显示直接接触共培养组c Tn T的表达均高于间接接触组共培养组(P<0.05)。结论直接和间接接触共培养模拟的心肌微环境均可诱导BMSCs向心肌样细胞分化,直接接触的分化率优于间接接触。

Objective To study the effect and the convert ratio comparison as well as the discussion of bone marrow mesenchymal stem cells( MSCs) co-cultured by different contact methods differentiating into cardiomyocyte-like cells.Methods By in vitro amplification of MSCs to study direct contact co-culture with the cardiomyocytes and simulated cardiac microenvironment of indirect contact co-culture established by transwell chambers,as well as their influence on differentiation of BMSCs into cardiomyocytes at the cellular level.Cell morphology was observed by inverted microscope.Two weeks later,the expression of the cardiac Troponin T( c Tn T),cardiac specific protein were detected by immunofluorescence and RT-PCR.Results Both immunofluorescence and RT-PCR showed that c Tn T expression by the direct contact co-cultural team was higher than that of indirect one( P < 0.05).Conclusions Both direct and indirect contact co-culture with simulated cardiac microenvironment could induce the MSCs to differentiate into cardiomyocyte-like cells,but the differentiation ratio of direct contact co-culture is higher than that of indirect contract co-culture.