血管外膜成纤维细胞诱导BMSCs向平滑肌细胞定向分化

Study on Differentiation of Bone Marrow Mesenchymal Stem Cells in Coculture with the Adventitial Fibroblasts Cells

观察大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)与血管外膜成纤维细胞(ad-ventitial fibroblasts,AF)直接接触培养后,BMSCs向血管成分细胞分化的情况.将BMSCs(DAPI标记)与外膜成纤维细胞按一定的比例混合培养7 d,BMSCs单独培养作对照,显微镜下观察细胞形态变化;用免疫荧光染色检测BMSCs的血管平滑肌细胞表型标志物肌动蛋白(SMα-actin)表达的情况;RT-PCR检测SMα-actin mRNA表达的情况.BMSCs与血管外膜成纤维细胞共培养后可见细胞核蓝染的BMSCs与SMα-actin表达阳性(红色)的双标细胞出现,且随培养时间的延长,BMSCs的SMα-actin表达阳性率增高.结果可见:与血管外膜成纤维细胞直接接触有诱导BMSCs向血管平滑肌细胞分化的趋势.

To assess the differentiation and morphological changes of bone marrow mesenchymal stem cells（BMSCs） as cocultured with adventitial fibroblasts cells.Rat BMSCs and adventitial fibroblasts cells were cocultured and evaluated.Firstly,BMSCs were labeled with DAPI,and then cocultured with adventitial fi-broblasts cells for 7 days.BMSCs were cultured alone as the control group.The changes of morphology of cocultured cells were demonstrated.Immunfluorescence analysis was performed by using antibodies against smooth muscle actin（SM α-actin）.The expression of SM α-actin mRNA was identified using RT-PCR.The BMSCs cocultured with adventitial fibroblasts cells expressed SM α-actin,the expression of SM α-actin or SM α-actin mRNA gradually increased with the extension of time.Results showed that cell-to-cell contact can induce rat BMSCs to differentiate into vascular smooth muscle like cells.