中文摘要：

为了探讨小鼠 MyoD 基因诱导绵羊脐带间充质干细胞（umbilical cord mesenchymal stem cells,UCMSCs）为成肌细胞的可能性,本研究用小鼠 MyoD-pcDNA3.1真核表达载体质粒转染绵羊 UCMSCs,在观察细胞形态变化的同时,检测成肌细胞标记蛋白表达、表达成肌细胞特异蛋白的细胞比率和成肌细胞特异基因 mRNA 相对表达量。结果显示,与对照组（未转染）相比,在转染 MyoD-pcDNA3.1质粒后第21天,大部分细胞呈现似成肌细胞的细长管状；与对照组未表达相关蛋白相比,转染 MyoD-pcDNA3.1后第8天,在荧光倒置显微镜下观察到细胞表达MyoD 和 Desmin 蛋白荧光,转染后第16天,不仅观察到细胞表达 MyoD 和 Desmin,而且观察到 MyoG 蛋白的表达；对于转染细胞后22 d 的细胞进行流式细胞仪检测显示,表达 MyoD、MyoG 和 Desmin 的细胞比率分别达93.5%、97.4%和99.5%；此外,实时荧光定量 PCR 检测显示,转染 MyoD-pcDNA3.1后第28天,其细胞中的MyoD、MyoG 和 Desmin mRNA 相对表达量分别提高2.046、2.389和5.489倍。上述结果表明,利用小鼠 MyoD构建真核表达载体具有诱导 UCMSCs 分化为成肌细胞的功效。

英文摘要：

In order to investigate the differentiation of sheep umbilical cord mesenchymal stem cells （UCMSCs）into muscle cells induced by mouse MyoD gene.This study based on the previ-ous work constructed the eukaryotic expression vector of MyoD-pcDNA3.1 in mice,and the vec-tor was transfected into sheep UCMSCs.The morphological changes of cells were observed by fluorescent microsco,the expression of MyoD ,Desmin and MyoG genes were detected by immu-nofluorescence,the percentage of cells expressing the cell specific factor （MyoD,Desmin and MyoG）was analyzed by flow cytometry and Real-time quantitative PCR to detect the relative ex-pression of mRNA relative to muscle cell specific factor.Compared with the control group （no transfection）,the vector was transfected into sheep UCMSCs,it was found that the cells were transformed into a long,slender,muscular cell state,and the cell spiral gradually disappeared at 21th day.It was found that MyoD and Desmin showed positive expression by immunofluorescence assay at 8th day,the expression of MyoG was also found after 1 6 d of induction,and the expres-sion of MyoD decreased,the amount of Desmin expression was no change;By flow cytometry,the percentages of the expression of MyoD,MyoG and Desmin were 93.5%,97.4% and 99.5%,respectively;Real-time quantitative PCR results showed that the relative expression of MyoD, MyoG and Desmin were increased and compared with the control group （non transfected cells）, the cells were increased by 2.046,2.389 and 5.489 times,respectively.The results showed that mouce MyoD gene could induce the differentiation of sheep UCMSCs into muscle cells.