Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (23): 4273-4278.doi: 10.3969/j.issn.1673-8225.2012.23.019

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In vitro proliferation of human bone marrow stromal cells after transfection by denovirus vector co-expressing human bone morphogenetic protein-2 and human fibroblast growth factor 2 genes

Guo Wei-tao1, Wang Hui1, Liu Si-jing2, Zeng Rong1, Xiao Qi-xian1, Chen Zi-qiu1, Huang Yun1, Wang Bin1, Hu Zi-bing1   

  1. 1Department of Orthopedics, Affiliated Hospital of Guandong Medical College, Zhanjiang 524001, Guangdong Province, China;
    2Affiliated Zhongshan Hospital of Guangzhou University of Traditional Chinese Medicine, Zhongshan 528400, Guangdong Province, China
  • Received:2011-11-03 Revised:2012-01-10 Online:2012-06-03 Published:2013-11-06
  • About author:Guo Wei-tao☆, M.D., Chief physician, Department of Orthopedics, Affiliated Hospital of Guandong Medical College, Zhanjiang 524001, Guangdong Province, China Guoweitao2000@sina.com

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Abstract:

BACKGROUND: Transfection of exogenous gene into tissue-engineered seed cells via recombinant adenovirus vector is the key to gene therapy of bone defects.
OBJECTIVE: To investigate the effect of gene transfection on the proliferation of human bone marrow stromal stem cells that tranfected with adenovirus vectors co-expressing human bone morphogenetic protein-2 (hBMP-2) and human fibroblast growth factor 2 (hFGF-2).
METHODS: The Ad-hBMP2-IRES-hFGF2 plasmids were transfected into human bone marrow stromal stem cells. The efficiency of transfection was evaluated by fluorescence microscope. Reverse transcriptase polymerase chain reaction was used to observe the successful transcription of hBMP-2 and hFGF-2 cNDA in bone marrow mesenchymal stem cells. Western blot assay was used to identify the protein expression of hBMP-2 and hFGF-2 genes. The cellular viability was determined by trypan blue staining and the changes of the cell proliferation were observed by flow cytometry assay.
RESULTS AND CONCLUSION: The expression of hBMP-2 and hFGF-2 genes was confirmed at mRNA and protein levels. The cell viability was not changed after transfection. The flow cytometry assay showed that the percentage of proliferating cells was increased significantly in cell cycle. The Ad-hBMP2-IRES-hFGF2 plasmids could transfect human bone marrow stromal stem cells with high transfection efficiency and promote cell proliferation.

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