Abstract:Objective:To investigate the method and effect of endothelial progenitor cells (EPCs) transfected with VEGF165 gene.
Methods :The cultivated EPCs were transfected with recombined adenovirus in which VEGF165 gene (Adv-GFP-VEGF165 group) or only Adv-GFP (Adv group) was incorporated,then the transfection effect was observed by immunofluorescence. And a blank control group was set up. Observation of the proliferation potential of cells was done by MTT method. The VEGF gene expression was observed by detecting the concentration of VEGF protein in supernatant with ABC-ELISA method.
Results:Green fluorescence was observed in almost all of the cells after transfection with Adv-GFP-VEGF165, which indicated the success of the transfection. The EPCs proliferation was not affected after the transfection, which was confirmed by MTT method. The VEGF protein concentration in the supernatant of transfected group was higher than that of Adv group and control group.
Conclusions: It is feasible for EPCs to be transfected with VEGF165 gene which was incorporated into adenovirus, and the suitable ratio was 1〖JP20〗∶〖JP〗50.The VEGF protein concentration in the supernatant liquid of endothelial progenitor cells transfected with VEGF increased significantly.