体外诱导人骨髓间充质干细胞向心肌细胞分化  

Inducing human mesenchymal stem cells into cardiomyocytes in vitro

作　　者：韩永生[1] 褚俊[1] 翟志敏[2] 严激[1] 周德存[3] 胡闻[4] 陈柯[4] 严中亚[3] 

机构地区：[1]安徽医科大学附属省立医院心内科,合肥230001 [2]安徽医科大学附属省立医院中心实验室,合肥230001 [3]安徽医科大学附属省立医院心外科,合肥230001 [4]安徽医科大学附属省立医院病理科,合肥230001

出　　处：《安徽医科大学学报》2005年第2期129-132,共4页Acta Universitatis Medicinalis Anhui

基　　金：安徽省优秀青年科技基金 (编号: 20020318 );安徽省教育厅自然科学基金(编号: 2004kj237zc)资助项目

摘　　要：目的　探索在体外诱导人骨髓间充质干细胞(hMSC)向心肌细胞分化的方法。方法　以5 杂氮胞苷( 5 Aza)对hMSC诱导,利用结蛋白抗体、房性利钠肽抗体以及闰盘蛋白抗体进行免疫组化染色,并进行超薄切片透射电镜观察。结果　经反复多次传代后,细胞形态及抗原表达无改变,始终保持未分化状态和稳定扩增。利用5 Aza对其诱导后第2周始,免疫组化染色发现细胞结蛋白、闰盘蛋白以及房性利钠肽表达均为阳性,不同浓度5 Aza诱导组之间差异无显著性。电镜观察发现这些细胞具有心肌细胞的特点。结论　在体外利用5 Aza可以诱导hMSC向心肌细胞分化。Objective To investigate the possibility of differentiation of human mesenchymal stem cells(hMSC) into cardiomyocytes.Methods hMSC were isolated by their adherence to plastic in vitro, then these cells were treated with 5-azacytidine for 24 h. After cultured for additional 2 weeks, these cells were identified by means of immunochemistry and transmission electron microscopy.Result hMSC showed a fibroblast- like morphology before treated with 5-azacytidine, but the morphology changed after 5-azacytidine treatment in most of the cells; they connected with adjoining cells after 1 week, formed myotube-like structures.2 weeks after induction, immunohistochemistry show that the induced cells were positive for desmin, connexin-43 and atrial natriuretic peptide. Electron microscopy revealed typical sarcomeres, a centrally positioned nucleus, and atrial granules-like ultrastructure.Conclusion This experiment proved that hMSC were able into cardiomyocyte after induction of 5-azacytidine in vitro .

关 键 词：骨髓祖代细胞/细胞学 骨髓细胞/药物作用 间质干细胞/细胞学 免疫组织化学 细胞分化/药物作用 显微镜检查 电子 心肌/细胞学 脱氧胞苷 

分 类 号：R322.11[医药卫生—人体解剖和组织胚胎学] R329.21[医药卫生—基础医学]