基础医学与临床 ›› 2010, Vol. 30 ›› Issue (6): 561-565.

• 研究论文 •    下一篇

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

赵艳梅 钟国强 柯红红 李金轶   

  1. 广西医科大学第一附属医院心内科 广西医科大学第一附属医院 广西医科大学第一附属医院心内科 广西医科大学第一附属医院
  • 收稿日期:2009-08-24 修回日期:2009-10-21 出版日期:2010-06-05 发布日期:2010-06-05
  • 通讯作者: 钟国强

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

Yan-mei ZHAO, Guo-qiang ZHONG, Hong-hong KE, Jin-yi LI   

  1. the First Affiliated Hospital, Guangxi Medical University the First Affiliated Hospital, Guangxi Medical University the First Affiliated Hospital, Guangxi Medical University
  • Received:2009-08-24 Revised:2009-10-21 Online:2010-06-05 Published:2010-06-05
  • Contact: Guo-qiang ZHONG,

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,第三代MSCs经DAPI标记后分别进行5-aza诱导,与CM共培养和单独培养。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。

关键词: 骨髓间充质干细胞, 心肌细胞, 5-氮胞苷, 共培养

Abstract: Objective After inducing and differentiating by different ways, we investigated the ability that mesenchymal stem cells( MSCs) differentiating into cardiomyocytes(CMs) and studied the differences that MSCs express 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 and cultured alone after labeled by 4 ,6-dimidino-2-phenylindone( DAPI) at the third passage. On the 1,2,3 and 4 weeks , DAPI-MSCs were observed the cell morphology and identified troponinT (cTnT) and connexin(Cx)43 by immunohistochemistry.Results DAPI-MSCs was't found 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 4 week. In the induction group ,the expression of cTnT and Cx43 was not observed at 1 week. In the co-culture group, the expressions of cTnT and Cx43 were detected at 5th day and increased with time .In the same weeks, 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 transformed into cardiomyocyte-like cells and expressed cTnT and Cx43. The ability of MSCs differentiated into CMs in the co-culture group was stronger than that of the induction group.

Key words: Bone marrow mesenchymal stem cells, cardiomyocytes, 5-Aza-2’-deoxycytidine, coculture