骨髓间充质干细胞的生物学特性及其向心肌样细胞的分化  

Biological Characteristics of Mesenchymal Stem Cells and Its Differentiation into Cardiocytes

作　　者：晋金兰[1] 庄汉屏[1] 

机构地区：[1]中南大学湘雅医院干部病房,湖南长沙410008

出　　处：《医学临床研究》2008年第4期594-597,共4页Journal of Clinical Research

基　　金：湖南省教育厅科学研究重点项目(湘财[2006]36号);湖南省卫生厅资助项目(B2006-038)

摘　　要：【目的】体外分离培养大鼠骨髓间充质干细胞(mesenchymal stem cells,MSCs),分析其生物学特性;5-氮胞苷(5-Azacytidine,5-aza)定向诱导其向心肌细胞分化。【方法】采用体外密度梯度离心法分离培养大鼠骨髓单个核细胞,流式细胞仪检测细胞表面标志;用含有5-aza(10μmol/L)的培养液培养第二代MSCs24h,诱导其向心肌细胞分化;诱导4周后采用免疫荧光技术检测心肌细胞特异性标志物Connexin 43和TroponinⅠ的表达。【结果】体外分离培养的细胞形态主要为纺锤形和长梭形,细胞表面抗原CD29、CD44表达为阳性,CD34、CD45表达为阴性;经5-aza诱导后细胞表达心肌细胞特异蛋白Connexin 43和TroponinⅠ。【结论】MSCs是存在于骨髓中的多能干细胞,体外经5-aza诱导可以定向分化为心肌样细胞。MSCs为心血管疾病的治疗提供了种子细胞。[Objective] To isolate and cultivate the mesenchymal stem cells （MSCs） from rat bone marrow, and to,analyze its biological characteristics and differentiation into cardiocytes induced by 5-Azacytidine （5-aza）. [Methods] The method of density gradient centrifugation was used to isolate rat bone marrow monocytes from neonate rat bone marrow. Surface antigen features of the MSCs were analyzed by flow cytometry. MSCs in second filial generation was treated by 5-aza （10μmol/L） for 24 hours for inducing its differentiation into cardiocytes. The specific proteins of cardiomyocytes, connexin 43 and troponin Ⅰ were detected by fluor-escence immunoassay at 4 weeks after treated by 5-aza. [Results] Rat bone marrow monocytes＇ shape mainly appeared as fusiform and long fusiform shape, and they were positive for CD29 and CD44, and negative for CD34 and CD45. After treated by 5-aza, they have been shown positive for specific proteins of cardiocytes, connexin 43 and troponin Ⅰ. [Conclusion] MSCs is a multipotent stem cell in the marrow, and can be differentiated into cardiocytes in vitro after treated by 5-aza. They provide and optimal stern cell source for therapy of cardiovascular disease.

关 键 词：骨髓细胞/细胞学 间质干细胞/细胞学 心肌/细胞学 

分 类 号：R329.21[医药卫生—人体解剖和组织胚胎学]