【摘 要】
：
目的 应用血小板衍生生长因子BB(platelet-derived growth factor BB,PDGF-BB),体外诱导人骨髓间充质干细胞(human bone marrow mesenchymal stem cells,hBMSCs)向血管平滑肌
【机 构】
：
上海第二医科大学附属第九人民医院整形外科,上海组织工程研究与开发中心
论文部分内容阅读
目的 应用血小板衍生生长因子BB(platelet-derived growth factor BB,PDGF-BB),体外诱导人骨髓间充质干细胞(human bone marrow mesenchymal stem cells,hBMSCs)向血管平滑肌细胞表型分化,探讨该方法的可行性及诱导细胞作为组织工程血管平滑肌种子细胞的可行性.方法 抽取健康成人志愿者骨髓,经密度梯度离心分离得单个核细胞,PDGF-BB(20 ng/ml)诱导hBMSCs向血管平滑肌样细胞(vascular smooth muscle cells,VSMCs)分化,观察细胞形态变化.免疫荧光检测细胞内血管平滑肌肌动蛋白α(vascular smooth muscle α-actin,SMα-actin),血管平滑肌钙结合蛋白(vascular smooth muscle calponin,SM calponin),血管平滑肌肌球蛋白重链(vascular smooth muscle myosin heavy chain,SMMHC)和细胞血管平滑肌钙结合相关蛋白(smooth muscle 22α,SM 22α)表达情况;反转录聚合酶链式反应(RT-PCR)检测诱导后血管平滑肌细胞SM α-actin,SM calponin,SMMHC和SM 22α的mRNA表达.Western印记检测SM 22α的表达.流式细胞分析技术(fluorescence activated cell sorter,FACS)分析诱导后细胞内SM α-actin,SM calponin,SMMHC的表达.结果 PDGF-BB 20ng/ml诱导后可见单层培养的细胞形态呈"成纤维细胞样".免疫荧光检测示SM α-actin,SM calponin,SMMHC,SM 22α表达阳性;RT-PCR检测SM α-actin,SM calponin,SMMHC,SM 22α的mRNA阳性表达.Western印迹检测SM 22α的表达为阳性.FACS分析表明诱导后,SMα-actin,SM calponin,SMMHC表达均增高,与未诱导组比较差异有统计学意义(P＜0.05,n=3).结论 人骨髓间充质干细胞在PDGF-BB的诱导下可向血管平滑肌细胞表型分化,有望成为血管组织工程血管平滑肌种子细胞的来源.
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