Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (40): 7441-7445.doi: 10.3969/j.issn.1673-8225.2011.40.008

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Enhanced green fluorescent protein gene-modified human bone marrow stromal cells induced by lentivirus

Tang Hao, Xiang Yong-sheng, Qin Ling-sha, Jiang Xiao-dan, Xu Ru-xiang, Chen Zhen-zhou   

  1. Department of Neurosurgery, Zhujiang Hospital, Southern Medical University, Guangzhou  510282, Guangdong Province, China
  • Received:2011-07-28 Revised:2011-09-07 Online:2011-10-01 Published:2011-10-01
  • Contact: Chen Zhen-zhou, Doctor, Associate chief physician, Department of Neurosurgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, Guangdong Province, China czz1020@163.com
  • About author:Tang Hao☆, Studying for doctorate, Department of Neurosurgery, Zhujiang Hospital, Southern Medical University, Guangzhou 　510282, Guangdong Province, China tanghao6251@gmail.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30801184*; the Medical Science Research Foundation of Guangdong Province, No. B2009159*; the Special Foundation for Science and Technology Key Plan of Guangdong Province, No. 2011A030400007*

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Abstract:

BACKGROUND: Transfection of primary culture cells is an important investigation field on cell biology, but the low transfection rate is one of the main obstacles which have limited its development and utility for a long time. Lentivirus transfection has become a good method because of its unique advantages in transfection of primary culture cells.
OBJECTIVE: To investigate the feasibility of transfection of human bone marrow stromal cells (hBMSCs) with lentivirus mediated enhanced green fluorescent protein (EGFP) gene.
METHODS: The hBMSCs were transfected by lentivirus carrying EGFP gene (LV-EGFP) at different MOI values under fetal bovine serum medium or serum-free condition respectively. The expressions of EGFP in these cells were observed for a long-term. These gene-modified cells were transplanted into the striatum rats of stereotaxicly. Survive of the grafted cells and EGFP expressions were detected by fluorescent microscope.
RESULTS AND CONCLUSION: After 48 hours of transfection, green fluorescent protein was successfully detected in hBMSCs. Transfection efficiency of cells in serum-free medium was higher than that in serum medium. The transfected hBMSCs could persistently express EGFP and had no obvious decline of fluorescence during 2-month culture in vitro. The EGFP positive cells could survive in the rat corpus striatum for more than 2 months and express EGFP persistently. Lentivirus is an efficient vector for gene modification of hBMSCs. Lentivirus mediated EGFP gene transfection can be used as efficient trance marker for hBMSCs transplantation study in vivo.

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