人脐血间充质干细胞诱导分化成心肌细胞的研究  

Human MSCs from UCB can differentiate into cardiomyocytes in vitro

作　　者：杨水祥[1] 黄景岭 王佐岩[1] 魏晓菲[1] 

机构地区：[1]北京大学医学部北京世纪坛医院心内科,北京100038 [2]青海省心血管病专科医院

出　　处：《中华老年心脑血管病杂志》2009年第4期292-295,共4页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

摘　　要：目的探讨体外脐血间充质干细胞诱导分化成心肌细胞的可行性和最佳方法。方法收集获知情同意的健康产妇脐血细胞,分离单个核细胞,从中进一步分离间充质干细胞,传代培养至第3代,应用免疫荧光流式细胞仪标记间充质干细胞特异性抗原CD34、CD44和CD90。5-氮胞苷诱导分化4周后,免疫组织化学染色和RT-PCR法分别检测心肌细胞标记物肌钙蛋白Ⅰ、转录因子GATA 4和β-肌球蛋白重链。结果在第3代细胞中,可检测到CD44、CD90的表达,未检测到CD34的表达。脐血间充质干细胞经5-氮胞苷诱导分化后,呈现成纤维细胞样形态和克隆增殖特点。免疫组织化学染色和RT-PCR可检测到肌钙蛋白Ⅰ,GATA 4和β-肌球蛋白重链的表达。结论脐血间充质干细胞能够被诱导分化成心肌样细胞,可成为干细胞移植的细胞来源。Objective To explore the possibility and best way of inducing mesenchymal stem cells （MSCs） from umbilical cord blood（UCB） to differentiate into cardiomyocytes in vitro. Methods The UCB was collected from the healthy delivery women with informed consent. The mononuclear cells in the blood were isolated, and MSCs were further separated and proliferated to the 3rd passage. The characteristic markers of MSCs,namely CD34,CD44 and CD90,were identified with the flow cytometry using immunofluorescence method. After inducing differentiation with 5-azacytidine for 4 weeks, the myocardial markers of troponin Ⅰ, transcription factor GATA 4 and β-myosin heavy chain（β-MHC） were examined by immunohistochemieal staining and RT-PCR respectively. Results The MSCs induced by 5-azacytidine showed fibroblast-like morphology and clonal proliferation characteristics. The immunophenotype of these cells was consistent with that reported for bone marrow mesenehymal stem cells. The myocardial markers of troponin I,GATA 4 and β-MHC were positively detected by immunohistochemical staining and RT-PCR respectively. Conclusion These data indicated that UCB can serve as a valuable and convenient source of mesenchymal stem cells, which can differentiate into cardiomyocyte-like cells, and can be as an alternative source in place of bone marrow-derived stem cells.

关 键 词：间质干细胞 胎血 肌钙蛋白Ⅰ 转录因子 肌球蛋白重链 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]