Transfection of VEGF gene in rabbit mesenchymal stem cells and its expression

Objective To explore the possibility of human vascular endothelial growth factor 165(hVEGF165) to transfect rabbit mesenchymal stem cells(MSCs),and establish the experimental foundation of angiogenesis tissue engineering organization and the treatment of ischemic disorders.Methods pcDNA3.0-VEGF165 eukaryotic expression vector was constructed,the vector was used directedly to transfect MSCs.The supernatant then was collected and the soluble protein of hVEGF gene expression was analysed with ELISA method.The proliferation capability of human umbilical vein endothelial cells(HUVEC) stimulated by the supernatant was measured with MTT methods,untreated MSCs and transfected MSCs were used as control groups.Results hVEGF eukaryotic expression vector was successfully constructed and transfected into the MSCs.Compared with the control group,concentration of hVEGF protein in the supernatant of the cultured cells increased significantly after the MSCs were tranfected with pcDNA3.0-VEGF165 for 24,48,72 h(P0.05),hVEGF protein was still found till the 12th progeny.The proliferation rate of HUVEC cells in the culture containing 2% supernatant of transfected cells was obviously higher than that of the control group(P0.05).Conclusion hVEGF gene could be successfully transfected into MSCs and could express effectively.