Experimental Study of Human Bone Marrow Mesenchymal Stem Cells Transfected with Vascular Endothelial Growth Factor Gene in Vitro

Abstract

Vascular endothelial growth factor 165 (VEGF165) is one of the most prominent vascular growth factors that has the potential to accelerate wound healing by promoting the regeneration of capillary vasculature. However, the short biological half-live of growth factors may impose severe restraints on their clinical applications. Here, we investigate the feasibility and safety of human bone marrow derived mesenchymal stem cells (MSCs) transfected with VEGF165 gene so as to provide a new method for clinical skin repair in the future. The MSCs were harvested, isolated and purified by combining density gradient centrifugation with adhering method. The pShuttle-CMV/VEGF165 was introduced into cultured MSCs through liposome-mediated transfection. The VEGF165 gene and protein expression in transfected MSCs and untransfected MSCs were determined by reverse transcription-polymerase chain reaction (RT-PCR), Enzyme-linked Immunoadsorbent Assay (ELISA) and western blot analysis respectively. We found that the expression of VEGF165 mRNA and protein in transfected MSCs were much higher than those in the untransfected MSCs. These results suggest that human bone marrow derived mesenchymal stem cells can be successfully transfected with vascular endothelial growth factor 165 gene mediated by liposome in vitro. The present study has laid a foundation for potential use of VEGF165 gene transfection to accelerate the vascularization of tissue engineered skins grafting.