microRNA定向诱导iPS细胞向心肌细胞分化的初步研究  

MiRNA Transfection Induces Oriented Differentiation of Induced Pluripotent Stem Cells into Cardiomyocyts

作　　者：吴仪 陈虎艳[1,2] 李宁 宋晓伟[3] 陈思锋[2] 袁文俊[1,3] 

机构地区：[1]宁夏医科大学基础医学院生理学系,银川750004 [2]复旦大学上海医学院生理与病理生理学系,上海200032 [3]第二军医大学生理学教研室,上海200433

出　　处：《宁夏医科大学学报》2012年第8期760-763,共4页Journal of Ningxia Medical University

基　　金：973计划项目(2009CB521902)

摘　　要：目的研究miRNAs诱导iPS细胞向心肌细胞的定向分化。方法将心肌细胞特异性miRNA-1、miR-NA-133a和miRNA-208排列组合后分别转染iPS细胞,悬浮培养3d后提取RNA和蛋白,应用Real-timePCR和Western blot的方法检测心肌细胞标志分子α-肌球蛋白重链(α-MHC)和肌钙蛋白T(Troponin-t)的表达。结果转染了miRNA-133a和miRNA-208+miRNA-133a组合的两个实验组中,α-MHC的mRNA表达量显著升高;同时转染了miRNA-208+miRNA-133a两个miRNA的实验组中,Troponin-t的mRNA和蛋白表达量显著上调。结论 miRNA-208和miRNA-133a共同作用,有利于诱导iPS细胞向心肌细胞的定向分化。Objective To explore oriented differentiation of induced pluripotent stem cells(iPSCs) into cardiomyocytes via miRNA transfection.Methods iPSCs were transfected with myocardial-specific miRNAs via permutation and combination of miRNA-1,miRNA-133a,miRNA-208.iPSCs were collected after 3 days suspension culture,from which RNA and protein were extracted for Real-time PCR and western blot analysis of α-MHC and Troponin-t,molecule markers of cardiomyocytes.Results Both α-MHC mRNA and protein levels increased significantly in iPSCs transfected with 'miRNA-133a' and 'miRNA-208 + miRNA-133a'.In addition,transfection with 'miRNA-208 + miRNA-133a'enhanced both mRNA and protein expressions of Troponin-t.Conclusion Co-transfection with miRNA-208 and miRNA-133a could induce oriented differentiation of iPSCs into cardiomyocytes.

关 键 词：IPS细胞 心肌细胞 MIRNA 

分 类 号：R541[医药卫生—心血管疾病]