体外不同诱导条件对大鼠骨髓间充质干细胞分化为心肌细胞的影响  

Effects of different conditions on rat bone marrow mesenchymal stem cells differentiating into cardiomyocytes in vitro

作　　者：赵艳梅[1] 钟国强[1] 柯红红[1] 李金轶[1] 

机构地区：[1]广西医科大学第一附属医院广西心血管病研究所心内科,广西南宁530021

出　　处：《基础医学与临床》2010年第6期561-565,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(30560051);广西留学回国人员科学基金(桂科回0448014);广西医疗卫生重点科研课题(桂卫重200301)

摘　　要：目的研究不同方法诱导骨髓间充质干细胞(MSCs)分化为心肌细胞(CM)的能力及表达心肌细胞特异性标记物的差别。方法分离培养MSCs及CM,第3代MSCs经DAPI标记后分别进行5-aza诱导、与CM共培养或单独培养。1、2、3和4周观察细胞形态变化及免疫细胞化学鉴定cTnT和Cx43。结果单独培养未见有细胞收缩,cTnT和Cx43表达阴性;诱导组和共培养组,培养至4周时细胞排列方向一致,成肌性排列,诱导1周时cTnT和Cx43表达阴性,而共培养组第5天cTnT和Cx43即开始表达,随着时间延长,两组cTnT和Cx43表达逐渐增加;相同周数内,共培养组的表达阳性率均高于诱导组(P<0.01)。结论 MSCs经5-aza诱导和与CM共培养,可转化为心肌样细胞并表达cTnT和Cx43,且共培养MSCs分化为心肌细胞的能力比5-aza诱导强。Objective To investigate the ability of mesenchymal stem cells（MSCs） differentiating into cardiomyocytes（CMs） and the differences that MSCs expression of cardiac cell-specific markers.Methods MSCs and CMs were isolated and cultivated,then MSCs were induced by 5-aza-2＇-deoxycytidine（5-aza）,cocultured with CMs or cultured alone after being labeled by 4,6-dimidino-2-phenylindone（DAPI） at the third passage.On week 1,2,3 and 4,DAPI-MSCs were observed for cell morphology and examined troponinT（cTnT） and connexin（Cx）43 by immunohistochemistry.Results DAPI-MSCs showed shrinkage and the expression of cTnT and Cx43 in the control group.In the induction group and the co-culture group,the cells were found to be orientating with one accord,arranging into a muscular at week 4.In the induction group,the expression of cTnT and Cx43 was not observed at week 1.In the co-culture group,the expressions of cTnT and Cx43 were found at 5th day and then increased.In the same period,the positive rate of cTnT and Cx43 in the co-culture group was higher than the rate of induction group（P〈0.01）.Conclusion After induced by 5-aza and co-cultured with CMs,the MSCs could transform intocardiomyocyte-like cells and expressed cTnT and Cx43.The ability of MSCs differentiating into CMs in the co-culture group was stronger than that of the induction group.

关 键 词：骨髓间充质干细胞 心肌细胞 5-氮胞苷 共培养 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]