天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (10): 1209-1212.doi: 10.11958/20160444

• 细胞与分子生物学 • 上一篇    下一篇

VEGF165在恒河猴骨髓间充质干细胞中的表达及鉴定

郭再玉， 张合亮， 水涛， 张国哲， 赵卫华， 陈谦， 侯延伟   

  1. 天津市泰达医院神经外科（邮编 300456）
  • 收稿日期:2016-05-20 修回日期:2016-08-30 出版日期:2016-10-15 发布日期:2016-10-21
  • 作者简介:郭再玉（1969）， 男， 副主任医师， 博士（后）， 主要从事神经外科研究
  • 基金资助:
    滨海新区卫生局科技项目（2011BHKZ007）

Expression and identification of VEGF165 in bone marrow mesenchymal stem cells of rhesus

GUO Zaiyu, ZHANG Heliang, SHUI Tao, ZHANG Guozhe, ZHAO Weihua, CHEN Qian, HOU Yanwei   

  1. Department of Neurosurgery, Teda Hospital of Tianjin, Tianjin 300456, China
  • Received:2016-05-20 Revised:2016-08-30 Published:2016-10-15 Online:2016-10-21

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摘要： 目的 探讨血管内皮生长因子（VEGF165）转染恒河猴间充质干细胞（MSCs）后的表达情况， 以及转染后对 MSCs 功能的影响。 方法 通过 Ficoll 法分离并培养恒河猴骨髓 MSCs， 进行细胞表型鉴定。 转染 pcDNA-eGFPVEGF165至 MSCs， 荧光显微镜观察增强型绿色荧光蛋白（eGFP）表达情况， 同时流式细胞技术对转染成功的细胞进行细胞表型及 eGFP 表达鉴定， RT-PCR 法检测转染后 VEGF165表达情况。 结果 成功分离到纯度较高的 MSCs， 转染后的 MSCs 及子代细胞均有 eGFP 和 VEGF165 的表达， 且保留了 MSCs 的特性。 结论 VEGF165 基因转染 MSCs 后可以稳定表达外源基因， 且可以维持 MSCs 的特性。

关键词: 间质干细胞, 血管内皮生长因子, 恒河猴, 转基因, 间质干细胞移植, 绿色荧光蛋白质类

Abstract: Objective To detect the transferred vascular endothelial growth factor (VEGF) 165 gene expression in rhesus autologous bone marrow mesenchymal stem cells (MSCs), and to explore the functional viability of transgenic MSCs. Methods MSCs from rhesus bone were isolated by Ficoll, which were used to detect the phenotype. After the culturing, the expression vector pcDNA- eGFP- VEGF165 was transfected into bone marrow MSCs. Fluorescence microscope and flow cytometry were used to detect the enhanced green fluorescent protein (eGFP) expression. At the same time, the phenotype in transfected MSCs was also indentified. The VEGF165 expression level was detected by RT- PCR. Results The highly purified MSCs were collected successfully. The transfected MSCs and daughter cells showed expressions of eGFP and VEGF165, which also remained the characteristics of MSCs. Conclusion The VEGF165 gene that is transfected into MSCs can maintain characteristics of MSCs, and stably express foreign genes.

Key words: mesenchymal stem cells, vascular endothelial growth factor, macaca mulatta, transgenes, mesenchymal stem cell transplantation, green fluorescent proteins