中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (14): 2520-2523.doi: 10.3969/j.issn.1673-8225.2012.14.010

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

三种大鼠骨髓间充质干细胞双向电泳样品制备方法的比较***★

覃万安，苏  伟，黄  钊，崔向荣，郭  杰   

  1. 广西医科大学第一附属医院创伤骨科手外科，广西壮族自治区南宁市  530021
  • 收稿日期:2011-09-19 修回日期:2011-10-26 出版日期:2012-04-01 发布日期:2012-04-01
  • 通讯作者: 苏伟，教授，主任医师，广西医科大学第一附属医院创伤骨科手外科，广西壮族自治区南宁市 530021 gxsuwei@163.com
  • 作者简介:覃万安★，男，1980年生，广西壮族自治区南宁市人，壮族，2004年广西右江民族医学院毕业，硕士，医师，主要从事骨髓间充质干细胞蛋白质组学的研究。qinwanangx@163.com
  • 基金资助:

    国家自然科学基金资助项目(30860078)；广西壮族自治区教育厅科研项目(200710MS021)及广西医科大学第一附属医院科研基金项目。

Three kinds of two-dimensional gel electrophoresis sample preparation methods of rabbit bone marrow mesenchymal stem cells  

Qin Wan-an, Su Wei, Huang Zhao, Cui Xiang-rong, Guo Jie   

  1. Branch of Hand Surgery, Department of Traumatic Orthopedics, First Affiliated Hospital of Guangxi Medical University, Nanning  530021,Guangxi Zhuang Autonomous Region, China
  • Received:2011-09-19 Revised:2011-10-26 Online:2012-04-01 Published:2012-04-01
  • Contact: author: Su Wei, Professor, Chief physician, Branch of Hand Surgery, Department of Traumatic Orthopedics, First Affiliated Hospital of Guangxi Medical University, Nanning 530021,Guangxi Zhuang Autonomous Region, China gxsuwei@163.com
  • About author:Qin Wan-an★, Master, Resident physician, Branch of Hand Surgery, Department of Traumatic Orthopedics, First Affiliated Hospital of Guangxi Medical University, Nanning 530021,Guangxi Zhuang Autonomous Region, China qinwanangx@163.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30860078*; Research Program of Education Department of Guangxi Zhuang Autonomous Region, No. 200710MS021*; Research Foundation of the First Affiliated Hospital of Guangxi Medical University*

PDF

344

摘要：

背景：骨髓间充质干细胞双向电泳样品制备方法很多，但目前尚未有统一标准。
目的：通过对比确定合适的大鼠骨髓间充质干细胞双向电泳样品制备方法。
方法：充分裂解体外培养的大鼠骨髓间充质干细胞，所得蛋白样品分为3组：未进一步处理组、丙酮沉淀法处理组、2-D clear-up kit处理组，Bradford法测定蛋白质浓度后分别进行双向电泳，对比分析3组所得电泳图谱。
结果与结论：2-D Clean-up kit处理组所得双向电泳图谱在背景，图像清晰度，分辨率，重复性上比丙酮沉淀法处理组和未进一步处理组要好，说明经2-D Clean-up kit处理的样品制备方法更适合用于大鼠骨髓间充质干细胞双向电泳样品的制备。
 

关键词: 双向电泳, 骨髓间充质干细胞, 大鼠, 蛋白质组学, 样品制备

Abstract:

BACKGROUND: A variety of “standard” sample preparation methods for two-dimensional gel electrophoresis have been reported, but there is no single one can be universally applied to rabbit bone marrow mesenchymal stem cells (rBMSCs).
OBJECTIVE: To identif a suitable sample preparation method for rBMSCs two-dimensional gel electrophoresis (2-DE).
METHODS: After full protein extraction of rBMSCs cultured in vitro, protein samples were divided into three groups: untreated, acetone precipitation and 2-D clean-up kit group, the protein concentration was measured by Bradford method, electrophoresis maps were analyzed.
RESULTS AND CONCLUSION: Using 2-D clean-up kit to treat the protein sample could obtain the best electrophoresis map which showed lowest background, clearest protein spots, highest resolution and good reproducibility. Protein extraction treated with 2-D Clean-up was a more suitable 2-DE sample preparation method of rBMSCs.

中图分类号: