Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (10): 1755-1758.doi: 10.3969/j.issn.1673-8225.2011.10.010

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Transient expression efficiency of foreign gene in human embryonic stem cells

Lu Jun, Chen Zong-feng, Huang Liang-hu, Tan Jian-ming   

  1. Fujian Provincial Key Laboratory of Transplant Biology, Fuzhou General Hospital of Nanjing Military Area Command of Chinese PLA, Fuzhou  350025, Fujian Province, China
  • Received:2010-11-28 Revised:2011-01-28 Online:2011-03-05 Published:2011-03-05
  • Contact: Tan Jian-ming, Doctoral supervisor, Professor, Fujian Provincial Key Laboratory of Transplant Biology, Fuzhou General Hospital of Nanjing Military Area Command of Chinese PLA, Fuzhou 350025, Fujian Province, China doctortjm@yahoo.com.cn
  • About author:Lu Jun☆, Doctor, Assistant researcher, Fujian Provincial Key Laboratory of Transplant Biology, Fuzhou General Hospital of Nanjing Military Area Command of Chinese PLA, Fuzhou 350025, Fujian Province, China junlu.heather@yahoo.com
  • Supported by:

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

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Abstract:

BACKGROUND: Technologies designed to allow modification and manipulation of human embryonic stem cells (hESCs) is numerous and various in the complexity of their efficiency, reliability and safety.
OBJECTIVE: To compare effect of different enhanced green fluorescent protein (EGFP) expression vectors with different promoter on expression efficiency in human embryonic stem cells (hESCs).
METHODS: The H9 cells cultured with non trophoblast were transfected with Fugene HD and lipofectamine 2000, positive colonies were detected under fluorescence microscope, and the expression efficiency were measured using flow cytometry analysis between different transfect vectors such as pCAG-EGFP and pEGFP-N3.
RESULTS AND CONCLUSION: Expression efficiencies of pCAG-EGFP and pEGFP-N3 were (42.45±3.32)% and (25.95±1.91)% respectively in Fugene HD transfected hESCs, and they had statistically difference (P < 0.05). It was (1.94±0.18)% and (1.49±0.33)% in lipofectamine 2000. The expression efficiencies of pCAG-EGFP and pEGFP-N3 transfected with Fugene HD were higher than that transfected with lipofectamine 2000 (P < 0.05).These results indicate that Fugene HD have the greater expression efficiency compared to lipofectamine 2000 in hESC line H9; expression efficiency of EGFP driven by CAG promoter is higher than that by CMV promoter.

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