《中国康复理论与实践》

《中国康复理论与实践》 ›› 2006, Vol. 12 ›› Issue (09): 783-784.

• 基础研究 • 上一篇    下一篇

成人脂肪间充质干细胞体外诱导分化为心肌细胞的实验研究

夏菁; 陈光辉; 刘宏斌; 王莉   

  1. 解放军总医院心内科,北京市 100853
  • 收稿日期:2006-03-30 出版日期:2006-09-01 发布日期:2006-09-01

Inducing Mesenchymal Stem Cells from Adult Human Adipose Tissue to Differentiate into Cardiomyocytes In Vitro

XIA Jing, CHEN Guang-hui, LIU Hong-bin, et al   

  1. Department of Cardiology, General Hospital of PLA, Beijing 100853, China
  • Received:2006-03-30 Published:2006-09-01 Online:2006-09-01

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被引次数

21

摘要： 目的体外培养并诱导成人脂肪间充质干细胞(ADMSCs)分化为心肌样细胞,为心肌细胞的替代治疗提供新的来源。方法采用消化法和贴壁培养法分离培养ADMSCs,对第3代细胞进行免疫化学鉴定和5氮杂胞苷(5aza)诱导,于诱导后第7、14、21和28天行免疫化学鉴定,并测定GATA4和Nkx2.5表达和心房利钠肽(ANP)含量。结果人脂肪中含有梭形细胞,CD44、CD13、CD105阳性表达,而CD45、CD34、HLADR、Ⅷ因子阴性表达。诱导后细胞排列方向一致,部分细胞聚集成团;结蛋白、α横纹肌肌动蛋白、肌球蛋白重链和心肌肌钙蛋白T呈阳性表达。有GATA4和Nkx2.5的表达及心房利钠肽的分泌。结论成人脂肪组织中含有间充质干细胞,且可在5氮杂胞苷诱导21d后分化为心肌样细胞。

关键词: 脂肪间充质干细胞, 心肌细胞, 转化, 体外诱导

Abstract: ObjectiveTo study the differentiation of adipose tissue-derived mesenchymal stem cells (ADMSCs) into cardiomyocytes in vitro. MethodsADMSCs were isolated and purified by the method of digesting and adhering to the culture plastis. The third generation of cells were determined with immunocytochemistry method and induced by 5-aza. On the 7th, 14th, 21st, 28th day after being induced, the surface antigen of myoblast cell and cardiac myocyte were determined, the expression of gene of GATA4 and Nkx2.5 were tested, and the content of atrial natriuretic polypeptide (ANP) assayed. ResultsImmunocytochemical staining showed CD44, CD13, CD105 positive, CD45, CD34, HLA-DR, factor Ⅷ negative. After being induced by 5-aza, the direction of cell arraying was gradually similar. The cells were stained positively for Desmin, α-Sarcomeric Actin, myosin heavy chain and Troponin T. The cells express GATA4 and Nkx2.5 and secrete ANP. ConclusionThere was mesenchymal stem cells in human adipose tissue. 5-aza may induce ADMSCs to differentiate into cardiomyocytes in vitro.

Key words: adipose tissue-derived mesenchymal stem cells, cardiomyocyte, differentiation, induce in vitro