CD47重组质粒及其过表达大鼠骨髓间充质干细胞的构建及鉴定

发布时间：2018-01-30 20:58

本文关键词： CD47 骨髓间充质干细胞重组质粒转染表达　出处：《重庆医科大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：构建CD47重组质粒并将其传染至大鼠骨髓间充质干细胞(bone mesenchymal stem cells, BMSCs)，建立稳定过表达CD47的BMSCs，为研究干细胞治疗中逃避免疫监视的有关机制提供实验模型。方法：构建过表达CD47基因的质粒载体，转染至分离培养的大鼠BMSCs，，RT-PCR、Western blot分别检测干细胞CD47mRNA、蛋白水平的表达，并完成功能检测。结果：成功构建了pYr-ads-4-ratCD47质粒，经酶切、测序结果表明pYr-ads-4-ratCD47重组质粒构建成功；RT-PCR、Western blot显示实验组转染至大鼠BMSCs后CD47的mRNA和蛋白水平表达明显增高（p0.05）；功能检测提示过表达CD47分子的BMSCs成功逃避巨噬细胞吞噬。结论：经质粒介导可成功将大鼠CD47目的基因片段转染至大鼠BMSCs，并能够在体外有效的过表达。
[Abstract]:Objective : To construct CD47 recombinant plasmid and transfer it to bone marrow mesenchymal stem cells ( MSCs ) in rat bone marrow mesenchymal stem cells ( MSCs ) , and to establish bone marrow cells stably expressing CD47 to provide experimental model for the study of immune surveillance in stem cell therapy . Methods : The expression of CD47mRNA and protein in stem cells was detected by RT - PCR and Western blot , and the function of CD47 mRNA was detected by RT - PCR and Western blot . Results : The expression of CD47 was successfully constructed by RT - PCR and Western blot , and the expression of CD47 mRNA and protein was significantly increased ( p < 0.05 ) . Conclusion : The gene fragment of rat CD47 can be successfully transfected into rat bone marrow cells via plasmid - mediated gene transfer , and can be effectively expressed in vitro .

【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R392

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