Experimental Study on Transfection of Adenovirus Containing VEGF Gene into Rat Mesenchymal Stem Cells in Vitro

[Objective] To investigate the expression of adenovirus transfected vascular endothelial growth factor (VEGF) in rat mesenchymal stem cells in vitro and the effect of transfection on MSCs proliferation. [Methods] A adenoviral vector containing VEGF165 gene was constructed and MSCs were isolated and expanded using the preplating method. The infection efficiency of adenovirus vector to MSCs was tested by Ad.GFP infection procedure. Ad.VEGF expression in MSCs and its secretion in culture medium were measured by immunohistochemical staining, Western blot, and ELISA, respectively. [Results] MSCs could be effectively transfected by adenovirus containing VEGF gene in vitro, the transfection efficiency has the dose-effect relationship with multiplicyties of infection (multiplicyties of infection, MOI). When MOI was 150, the infection efficiency was more than 95%. The expression of VEGF was traced both in cell lysate and in culture medium. A maximum production of VEGF was observed at 3～9 days after infection (1 125 pg/mL at the ninth day), and VEGF was found even in day 13. [Conclusion] Gene transfer technology mediated by adenoviral vector can transfect VEGF gene into MSCs with high efficiency. MSCs transfected by VEGF gene could express VEGF.