c-Met基因过表达脐带间充质干细胞株的构建  

Construction of overexpression of c-met in human umbilical cord mesenchymal stem cells with lenti-vector transfection in vitro

作　　者：张永婷 朱传龙 李军[1] 章莉莉 Zhang Yongting;Zhu Chuanlong;Li Jun;et al(Department of InfectiousDisease, First Affiliated Hospital, Nanjing Medical University, Nanjing 210029, Jiangsu Province, China)

机构地区：[1]南京医科大学第一附属医院感染病科,南京市210029

出　　处：《实用肝脏病杂志》2018年第3期397-400,共4页Journal of Practical Hepatology

基　　金：国家自然科学基金资助项目(编号:81770591);江苏省高层次卫生人才"六个一工程"资助项目(编号:LGY2016005)

摘　　要：目的建立c-Met慢病毒载体转染人脐带间充质干细胞(hUMSC),为治疗肝衰竭作为种子细胞。方法培养hUMSC,经流式细胞技术检测细胞表面表型,转染c-Met慢病毒载体,在荧光显微镜下观察转染效率,确定最佳多重感染复数(MOI)。采用嘌呤霉素抗性筛选稳定表达c-Met的hUMSC细胞系,采用Western-blot法检测细胞c-met蛋白表达量。结果 P5代细胞高表达CD44、CD90和CD105抗原,不表达CD31、CD45和CD34相关造血细胞抗原,符合人脐带间充质干细胞的特性;构建成功的c-Met慢病毒载体转染人脐带间充质干细胞最佳MOI=80,经嘌呤霉素筛药,在荧光显微镜下观察发现荧光阳性率为100%,且经Western-blot法检测证实该细胞过表达c-Met蛋白。结论成功构建过表达c-Met基因的脐带间充质干细胞,为进一步实验打下了基础。Objective To construct the overexpression of c-met in human umbilical cord mesenchymal stemcells（hUMSCs）with lenti-vector transfection in vitro. Methods The phenotype of hUMSCs was determined byFlow cytometry analysis,and the hUMSCs were transfected by different multiple of infection（MOI with c-metlenti-vector. The optimal MOI was determined under fluorescence microscopy. The c-Met protein was detected byWestern blot. Results P5 gegeration of hUMSCs were CD44/CD90/CD105 -positive, and CD31/CD45/CD34-negative,indicating the hUMSCs were obtained with specific surface markers. The optimal MOI was 80,and afterpuromycin screening, the transfected efficiency was up to 100%. The c -Met protein was overexpressed in thetransfected hUMSCs by Western blot detection. Conclusion c -Met overexpressing hUMSCs were successfullyconstructed,which might be beneficial for further study.

关 键 词：人脐带间充质干细胞 C-MET基因 转染 体外 

分 类 号：R575.3[医药卫生—消化系统]