中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (27): 5017-5021.doi: 10.3969/j.issn.2095-4344.2012.27.015

• 干细胞转基因表达 transgenic expression in stem cells • 上一篇    下一篇

pIRES-骨形态发生蛋白2质粒转染大鼠骨髓间充质干细胞后的 持续表达

孙 立1，田晓滨1，胡如印1，王远政1，田家亮1，韩 伟1，李 波1，袁 军2，聂瑛洁2   

  1. 贵州省人民医院，1骨科， 2中心实验室，贵州省贵阳市 550002
  • 收稿日期:2012-04-19 修回日期:2012-05-10 出版日期:2012-07-01 发布日期:2013-11-01
  • 通讯作者: 田晓滨，主任医师，贵州省人民医院骨科，贵州省贵阳市 550002 txb6@vip.163.com
  • 作者简介:孙立☆，1973年生，贵州省安顺市人，汉族，2007年同济医学院毕业，博士，副主任医师，主要从事骨组织工程、关节外科的研究。 sunly111@yahoo.cn
  • 基金资助:

    国家自然科学基金项目(81060145)；贵州省攻关项目(SY[2010]3057)；贵州省省长资金项目[(2008)100]；贵州省卫生厅基金项目(2008-1-019)。

Persistent expression of pIRES-bone morphogenetic protein 2 plasmid after transfection into rat bone marrow mesenchymal stem cells

Sun Li1, Tian Xiao-bin1, Hu Ru-yin1, Wang Yuan-zheng1, Tian Jia-liang1, Han Wei1, Li Bo1, Yuan Jun2, Nie Ying-jie2   

  1. 1Department of Orthopedics, 2Central Laboratory, People’s Hospital of Guizhou Province, Guiyang 550002, Guizhou Province, China
  • Received:2012-04-19 Revised:2012-05-10 Online:2012-07-01 Published:2013-11-01
  • Contact: Tian Xiao-bin, Chief physician, Department of Orthopedics, People’s Hospital of Guizhou Province, Guiyang 550002, Guizhou Province, China txb6@vip.163.com

PDF

397

被引次数

16

摘要：

背景：重组人骨形态发生蛋白2已被广泛应用于骨组织工程治疗骨缺损或骨不连，但是直接应用外源性骨形态发生蛋白2修复骨缺损效果不理想。
目的：观察转染pIRES-骨形态发生蛋白2的大鼠骨髓间充质干细胞持续合成并分泌骨形态发生蛋白2的情况。
方法：全骨髓贴壁法分离培养大鼠骨髓间充质干细胞，脂质体介导下将真核表达质粒pIRES-骨形态发生蛋白2导入大鼠骨髓间充质干细胞。
结果与结论：ELISA结果显示，随着pIRES-骨形态发生蛋白2转染时间的延长，大鼠骨髓间充质干细胞分泌的骨形态发生蛋白2逐渐增多，至转染后10~12 d达高峰，转染后15 d，仍可见较多骨形态发生蛋白2的表达。说明转染pIRES-骨形态发生蛋白2的大鼠骨髓间充质干细胞可持续稳定分泌骨形态发生蛋白2。

关键词: 骨形态发生蛋白2, 骨髓间充质干细胞, 细胞转染, 表达, 组织工程, 干细胞

Abstract:

BACKGROUND: Recombinant human bone morphogenetic protein has been widely used in bone tissue engineering for treatment of bone defects or bon nonunion. But direct application of exogenous bone morphogenetic protein for repair of bone defects yields unsatisfactory effects.
OBJECTIVE: To investigate persistent secretion and expression of bone morphogenetic protein in rat bone marrow mesenchymal stem cells transfected with pIRES-bone morphogenetic protein.
METHODS: Rat bone marrow mesenchymal stem cells were isolated and cultured by the whole bone marrow adherence method. Under the liposome induction, eukaryotic expression plasmid pIRES-bone morphogenic protein 2 was introduced into rat bone marrow mesenchymal stem cells.
RESULTS AND CONCLUSION: ELISA results showed that with the prolonged transfection time of pIRES-bone morphogenetic protein, bone morphogenetic protein 2 secreted by rat bone marrow mesenchymal stem cells was gradually increased, peaked at 10-12 days after transfection, and was relatively much at 15 days after transfection. These findings suggest that after transfected by pIRES-bone morphogenetic protein 2, rat bone marrow mesenchymal stem cells can persistently and stably secrete bone morphogenetic protein 2.

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