microRNA-133诱导绵羊脐带间充质干细胞分化为成肌细胞的研究  

Study on the differentiation of sheep umbilical cord mesenchymal stem cells into myoblasts induced by microRNA-133

作　　者：杜文敬 宋维文 陈胜男 朴善花 王春生 安铁洙 DU Wenjing;SONG Weiwen;CHEN Shengnan;PIAO Shanhua;WANG Chunsheng;AN Tiezhu(College of Life Science,Northeast Forestry University,Harbin 150040,China)

机构地区：[1]东北林业大学生命科学学院

出　　处：《黑龙江畜牧兽医》2018年第23期10-12,264,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：中央高校基本科研业务费专项(DL13EA06-02);东北林业大学大学生创新创业训练计划项目(201710225305)

摘　　要：为了探明microRNA-133(miR-133)在诱导脐带间充质干细胞分化为成肌细胞过程中的作用,试验采用脂质体法以化学合成的miR-133转染绵羊脐带间充质干细胞并进行续培养。当细胞呈现成肌细胞的形态特征时,采用qRT-PCR法、免疫荧光法和流式细胞术分别检测转染细胞中成肌细胞特异性基因Desmin与Myf5mRNA的相对表达量、成肌细胞标记蛋白Desmin与MyoD的表达情况及表达成肌细胞标记蛋白的细胞比率。结果表明:转染并续培养后第28天,续培养的细胞有序生长并呈现融合趋势;qRT-PCR检测可见,转染细胞中成肌细胞特异性基因Desmin及Myf5mRNA的相对表达量明显提高;与未转染细胞相比,转染细胞特异性表达成肌细胞标记蛋白MyoD和Desmin;此外,转染细胞中表达MyoD和Desmin的细胞比率分别为99.3%和99.0%。说明miR-133在绵羊脐带间充质干细胞分化为成肌细胞过程中发挥重要作用。The aim of the present study was to investigate the function of microRNA-133 ( miR-133) in inducing umbilical cord mesenchymal stem cells to differentiate into myoblasts. The sheep umbilical cord mesenchymal stem cells were transfected with chemically synthesized miR- 133 by using liposome method and then cultured. When the cells take on the morphological characteristics of myoblast,the relative expression levels of myoblast specific gene Desmin and Myf5 mRNA,the expression of myoblast marker protein Desmin and MyoD and percentage of cells expressing myoblast marker proteins were detected by qRT-PCR,immunofluorescence and flow cytometry,respectively. The results showed that on the 28th day after transfection and continuous culture,the cells grew in an orderly manner and showed a tendency of fusion. The relative expression levels of myoblast specific genes Desmin and Myf5 mRNA in transfected cells were significantly increased by qRT-PCR. Compared with untransfected cells,transfected cells specifically expressed myoblast marker proteins MyoD and Desmin. In addition,the percentage of transfected cells expressing MyoD and Desmin was 99. 3% and 99. 0%,respectively. These results suggest that miR-133 plays an important role in the differentiation of sheep umbilical cord mesenchymal stem cells into myoblasts.

关 键 词：miR-133 绵羊 间充质干细胞 诱导分化 成肌细胞 

分 类 号：S826[农业科学—畜牧学]