Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (10): 1769-1772.doi: 10.3969/j.issn.1673-8225.2011.10.013

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Protein expression of bone marrow mesenchymal stem cells after human vascular endothelial growth factor 165 gene transfection

Tian Li1, Liang Xiao-peng2, Tian Xiao-ye3, Yu Xin-chen4, Tian Jing4   

  1. 1Department of Clinic, Shenyang Medical College, Shenyang  110034, Liaoning Province, China; 2School of Optometry, Shenyang Medical College, Shenyang  110034, Liaoning Province, China; 3Jingkang Rehabilitation Center for Aged, Tieling Economic Development Zone, Tieling  112000, Liaoning Province, China; 4Shenyang Jingkang Medical Group, Shenyang  110043, Liaoning Province, China
  • Received:2010-10-03 Revised:2011-01-10 Online:2011-03-05 Published:2011-03-05
  • About author:Tian Li, Professor, Chief physician, Department of Clinic, Shenyang Medical College, Shenyang 110034, Liaoning Province, China Linan1956@yahoo.com.cn
  • Supported by:

    Educational Commission of Liaoning Province, No. 2008-7-29*; Foundation of Shenyang Technology Bureau, No. 1081233-1-00-03*

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Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) is the best source of seed cells for tissue engineering, which containing vascular endothelial growth factor 165 (VEGF165) not only has an important significance in angiogenesis and the start of bone repair, but also its sustained release can increase the degree of mineralization of new bone, and enhance the mechanical properties of repair tissue.
OBJECTIVE: To observe the hVEGF165 protein function expressed by the gene transfected BMSCs.
METHODS: BMSCs from rabbits were isolated and cultured in vitro, and which were purified and identified. Cell surface makers were detected by immunofluorescence assay. And the cultured BMSCs were transfected with the composite of PcDNA3.1-hVEGF165 plasmid and lipofectamine in the ratio of 1/3 mixed liquor, and which were divided into 3 groups: transfection group, empty vector transfection group, and non-transfection group. PcDNA3.1-hVEGF165 plasmid was used to transfect cells in transfection group; pcDNA3.1-empty vector transfection was applied in empty vector transfection group; no treatment was performed in non-transfection group. The expression of exogenous hVEGF165 was detected by ELISA and Western-blot.
RESULTS AND CONCLUSION: Compared with empty vector transfection group and non-transfection group, the protein level of VEGF165 in transfection group was significantly higher (P < 0.05), however, there was no significant difference between empty vector transfection group and non-transfection group (P > 0.05). There were significant differences of the protein level of VEGF165 in transfection group (P < 0.05) at different time points. The transfected BMSCs by hVEGF165 can successfully secret the protein of VEGF165. It is indicated that hVEGF165 can be transfected to BMSCs and efficiently express VEGF165 protein of biological activity by using of gene transfection technology.

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