中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (40): 7421-7424.doi: 10.3969/j.issn.1673-8225.2011.40.003

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

脂质体介导骨保护素基因转染大鼠骨髓基质干细胞及其表达

赵济华1，李琪佳1，郭  静1，王志强2   

  1. 河北联合大学，1实验中心，2 附属医院骨科，河北省唐山市063000
  • 收稿日期:2011-04-11 修回日期:2011-06-22 出版日期:2011-10-01 发布日期:2011-10-01
  • 通讯作者: 李琪佳，教授，河北联合大学实验中心，河北省唐山市063000 qjl1222@hotmail.com
  • 作者简介:赵济华★，女，1981年生，河北省唐山市人，河北联合大学在读硕士，主要从事骨组织工程方面的研究。 wxjhxz@sina.com
  • 基金资助:

    河北省人事厅留学归国人员科技活动基金资助(200911)。

Expression of osteoprotegerin gene mediated with liposome in rat bone marrow stromal cells

Zhao Ji-hua1, Li Qi-jia1, Guo Jing1, Wang Zhi-qiang2   

  1. 1Laboratory Center, Hebei United University, Tangshan  063000, Hebei Province, China; 2Department of Orthopaedics, Affiliated Hospital of Hebei United University, Tangshan  063000, Hebei Province, China
  • Received:2011-04-11 Revised:2011-06-22 Online:2011-10-01 Published:2011-10-01
  • Contact: Li Qi-jia, Professor, Laboratory Center, Hebei United University, Tangshan 063000, Hebei Province, China qjl1222@hotmail.com
  • About author:Zhao Ji-hua★, Studying for master’s degree, Laboratory Center, Hebei United University, Tangshan 063000, Hebei Province, China wxjhxz@sina.com
  • Supported by:

    the Foundation of Department of Human Resources of Hebei Province, No. 200911*

PDF

252

被引次数

3

摘要：

背景：基因修饰骨组织工程种子细胞，可提高对骨缺损的修复能力。
目的：构建含有骨保护素的真核表达载体并检测转染骨髓基质干细胞后的表达。
方法：取4周龄SD大鼠胫骨和股骨行骨髓基质干细胞的分离和培养，分为载体组、空载体组、对照组。空载体组转染plRES2-EGFP载体，载体组转染plRES2-EGFP-OPG。
结果与结论：转染后激光扫描共聚焦显微镜观察可见骨髓基质干细胞内绿色荧光蛋白表达；RT-PCR检测结果可见质粒载体组在1 200 bp有明显条带，空载体组及对照组未见表达；免疫细胞化学检测骨髓基质干细胞内骨保护素蛋白呈阳性；MTT法检测各组细胞增殖活力无明显改变(P > 0.05)。证实应用plRES2-EGFP-骨保护素质粒转染大鼠骨髓基质干细胞，可获得外源性骨保护素的基因及蛋白表达。

关键词: 脂质体, 骨髓基质干细胞, 骨保护素, 基因转染, 增殖活力

Abstract:

BACKGROUND: Genetically modified seed cells for bone tissue engineering can improve the ability to repair bone defects.
OBJECTIVE: To detect the expression of osteoprotegerin (OPG) in rat transfected bone marrow stromal cells (BMSCs).
METHODS: BMSCs were aseptically obtained from the femur and tibia of 4-week-old SD rats and cultured in vitro. BMSCs were divided into three groups: plasmid group, blank plasmid group and non-transfection group.
RESULTS AND CONCLUSION: After transfection, green fluorescent protein was observed in BMSCs under the laser scanning confocal microscope. There was obvious mark at 1 200 bp in plasmid group, but there was no expression of OPG mRNA in blank plasmid group and non-transfection group. The OPG protein was positive in BMSCs detected by immunocytochemistry examination. There were no significant difference in cell proliferation in each group (P > 0.05). plRES2- EGFP-OPG plasmid transfected BMSCs can express exogenous OPG and proteins. 

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