脂肪间充质干细胞体外分化成心肌样细胞  

In Vitro Differentiation of Rat Adipose-derived Stem Cells into Cardiomyocytes

作　　者：王二云[1] 赵丽丽[1] 高倩[1] 杨秋梅[1] 杨公社[1] 

机构地区：[1]西北农林科技大学动物脂肪沉积与肌肉发育实验室,陕西杨凌712100

出　　处：《中国生物化学与分子生物学报》2009年第9期855-860,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家重点基础研究发展计划(973计划; No.2004CB117506)~~

摘　　要：探讨大鼠脂肪间充质干细胞(adipose-derived mesenchymal stem cells,AMSC)体外分化成心肌样细胞的潜能,为自体干细胞移植治疗心肌梗死提供理论基础.采用消化法分离大鼠AMSC,培养于RPMI1640生长培养基中,倒置相差显微镜观察细胞形态发现,随着培养时间的延长,细胞形态趋向于心肌细胞,SQRT-PCR检测表达心肌特异性基因:β-肌球蛋白重链(β-MHC)、α-肌球蛋白重链(α-MHC)、心房利钠肽(ANP)、心肌肌钙蛋白(cTnT)、心肌肌动蛋白、肌肉增强因子和GATA-4;免疫细胞化学和免疫荧光染色检测表达心肌细胞特异性蛋白:结蛋白、横纹肌辅肌动蛋白、心肌肌动蛋白和间隙连接蛋白45(connexin45);Western印迹检测表达心肌特异性蛋白Nkx2.5.实验表明,大鼠AMSC在体外培养条件下能分化成心肌样细胞,在组织工程学及干细胞移植领域有着良好的应用前景.The potential to differentiate into cardiomyocyte from rat adipose-derived mesenchymal stem cells（AMSCs） was investigated,AMSCs were isolated from SD rat adipose tissue by digestion and maintained in the RPMI1640 medium.AMSCs transformed into cardiomyocyte-like cells during the long-term culture.The expression of β-MHC,ANP,cTnT,GATA-4,α-cardiac actin and MEF2C were detected by Quantitative RT-PCR.Cardiac-specific proteins,such as desmin,sarcomeric,α-actinin,α-cardiac actin and connexin 45 were detected by immuocytochemistry or immunofluorescence staining. The expression of early cardiac- specific protein Nkx2.5 were detected by Western blot. The results showed that rat AMDCs was able to differentiate into a cardiomyocyte in vitro, which could be used as a source for tissue engineering or stem cell transplantation for the treatment of ischemic myocardial infarction.

关 键 词：脂肪间充质干细胞(AMSC) 心肌样细胞 自发分化 

分 类 号：Q254[生物学—细胞生物学]