中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (49): 9151-9154.doi: 10.3969/j.issn.1673-8225.2011.49.007

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

新型化学转染试剂对人脐带间充质干细胞的转染优化

路  君，赵红州，陈  津，付云峰，黄梁浒，谭建明   

  1. 解放军南京军区福州总医院福建省移植生物学重点实验室，福建省福州市  350025
  • 收稿日期:2011-05-25 修回日期:2011-07-20 出版日期:2011-12-03 发布日期:2011-12-03
  • 通讯作者: 谭建明，博士生导师，教授，解放军南京军区福州总医院福建省移植生物学重点实验室，福建省福州市350025 doctortjm@yahoo.com.cn
  • 作者简介:路君☆，女，1981年生，福建省福州市人，汉族，2008年复旦大学毕业，博士，主管技师，主要从事干细胞分化研究。 junlu.heather@yahoo.com
  • 基金资助:

    福建省自然科学基金项目(2010J05078，2010J01217)。

Optimizing transfection of human umbilical cord-derived mesenchymal stem cells using novel chemical transfection reagents

Lu Jun, Zhao Hong-zhou, Chen Jin, Fu Yun-feng, Huang Liang-hu, Tan Jian-ming   

  1. Fujian Provincial Key Laboratory of Transplant Biology, Fuzhou General Hospital of Nanjing Military Region of Chinese PLA, Fuzhou  350025, Fujian Province, China
  • Received:2011-05-25 Revised:2011-07-20 Online:2011-12-03 Published:2011-12-03
  • Contact: Tan Jian-ming, Doctoral supervisor, Professor, Fujian Provincial Key Laboratory of Transplant Biology, Fuzhou General Hospital of Nanjing Military Region of Chinese PLA, Fuzhou 350025, Fujian Province, China doctortjm@yahoo.com.cn
  • About author:Lu Jun☆, Doctor, Technician in charge, Fujian Provincial Key Laboratory of Transplant Biology, Fuzhou General Hospital of Nanjing Military Region of Chinese PLA, Fuzhou 350025, Fujian Province, China junlu.heather@yahoo.com
  • Supported by:

    the Natural Science Foundation of Fujian Province, No. 2010J05078*, 2010J01217*

PDF

501

被引次数

4

摘要：

背景：人脐带间充质干细胞的基因修饰技术多种多样，它们在效率、可靠性及安全性方面各有不同。
目的：对比3种新型化学转染试剂在人脐带间充质干细胞中的瞬时转染效率。
方法：采用Fugene HD、Lipofectamine LTX及Attractene分别转染人脐带间充质干细胞，荧光显微镜下观察阳性细胞，流式细胞术分析不同转染试剂的转染效率，锥虫蓝染色观察转染后人脐带间充质干细胞的生存率。
结果与结论：Lipofectamine LTX转染效率最高，显著高于Fugene HD和Attractene[(32.50±2.12)%，(4.30±0.64)%，(1.70±0.08)%，P < 0.05]。Lipofectamine LTX转染后的细胞生存率略低于Fugene HD及Attractene，但差异无显著性意义[(69.8±6.3)%，(92.4±4.2)%，(106.6±3.9)%，P > 0.05]。提示Lipofectamine LTX是人脐带间充质干细胞较为理想的化学转染试剂。

关键词: 化学转染试剂, 脐带间充质干细胞, 基因转染, 表达效率, 安全性

Abstract:

BACKGROUND: Genetic modification technology of human umbilical cord-derived mesenchymal stem cells (hUCMSCs) is numerous and varies in the complexity of their efficiency, reliability and safety.
OBJECTIVE: To compare transfected efficiency of different novel chemical transfection reagents on hUCMSCs.
METHODS: The hUCMSCs were transfected with Fugene HD, Lipofectamine LTX, and Attractene. Positive cells were detected under fluorescent microscope, and expression efficiencies were measured by using flow cytometry. Cell viability was measured by using trypan blue exclusion after transfection.
RESULTS AND CONCLUSION: The highest transfected efficiency was observed with Lipofectamine LTX (32.5±2.12)% (P < 0.05), followed by Fugene HD (4.3±0.64)%, minimal transfection efficiency was observed with Attractene (1.74±0.08)%. Lipofectamine LTX demonstrated the lower cell viability (69.8±6.3)% compared with Fugene HD (92.4±4.2)% and Attractene (106.6±3.9)% (P > 0.05), but the difference was not significant. In summary, this study demonstrates that Lipofectamine LTX is a relative ideal chemical transfection reagent for hUCMSCs.

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