| c-Met基因过表达脐带间充质干细胞株的构建* |
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| 引用本文： | 张永婷,朱传龙,李军,章莉莉.c-Met基因过表达脐带间充质干细胞株的构建*[J].实用肝脏病杂志,2018,21(3):397-400. |
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| 作者姓名： | 张永婷 朱传龙 李军 章莉莉 |
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| 作者单位： | 210029 南京市 南京医科大学第一附属医院感染病科 |
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| 基金项目： | *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） |
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| 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。
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| 关 键 词： | 人脐带间充质干细胞 c-Met基因 转染 体外 |
| 收稿时间： | 2017-10-09 |
Construction of overexpression of c-met in human umbilical cord mesenchymal stem cells with lenti-vector transfection in vitro |
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| Zhang Yongting,Zhu Chuanlong,Li Jun,et al.Construction of overexpression of c-met in human umbilical cord mesenchymal stem cells with lenti-vector transfection in vitro[J].Journal of Clinical Hepatology,2018,21(3):397-400. |
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| Authors: | Zhang Yongting Zhu Chuanlong Li Jun |
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| Affiliation: | Department of Infectious Disease,First Affiliated Hospital,Nanjing Medical University,Nanjing 210029,Jiangsu Province,China |
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| Abstract: | Objective To construct the overexpression of c-met in human umbilical cord mesenchymal stem cells（hUMSCs） with lenti-vector transfection in vitro. Methods The phenotype of hUMSCs was determined by Flow cytometry analysis, and the hUMSCs were transfected by different multiple of infection（MOI with c-met lenti-vector. The optimal MOI was determined under fluorescence microscopy. The c-Met protein was detected by Western 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 after puromycin screening,the transfected efficiency was up to 100%. The c-Met protein was overexpressed in the transfected hUMSCs by Western blot detection. Conclusion c-Met overexpressing hUMSCs were successfully constructed, which might be beneficial for further study. |
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| Keywords: | Human umbilical cord mesenchymal stem cells c-Met Transduction In vitro |
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