血管外膜成纤维细胞诱导BMSCs向平滑肌细胞定向分化

摘要

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 co-cultured and evaluated. Firstly, BMSCs were labeled with DAPI, and then cocultured with adventitial fibroblasts 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 a-actin). The expression of SM a-actin mRNA was identified using RT-PCR. The BMSCs cocultured with adventitial fibroblasts cells expressed SM a-actin, the expression of SM a-actin or SM a-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.%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(adventitial 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7d,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(SM α-actin)表达的情况;RT-PCR检测SM α-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SM α-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SM α-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.