Study of adenovirus-mediated vascular endothelial growth factor 165 gene transfer into bone marrow stromal cells and its expressive product promoting proliferation of endothelial cells

Abstract

To study transcription, expression, and bioactivity of expressive protein of vascular endothelial growth factor 165 (VEGF(165)) gene, after bone marrow stromal cells (BMSCs) were transferred by VEGF(165) gene mediated by adenovirus vector ex vivo. BMSCs were Isolated and cultured by gradient centrifugation method. The cells cultured are transferred by recombinant adenovirus vector that carry VEGF(165) gene. Transcription and expression of VEGF(165) gene in BMSCs and secretion of VEGF protein in culture medium were measured by reverse transcriptase-polymerase chain reaction (RT-PCR), Western blot and enzyme linked immunosorbent assay (ELISA) methods. The activity of VEGF protein in culture media was detected by proliferation effects on vascular endothelial cells. When Ad. VEGF(165) transferred into BMSCs, there was an effective transcription and expression. The expressed product in the media of transferred cells had highly biological activity on proliferation of rat aortic vascular endothelial cells (P < 0.01). Adenovirus vector can be transferred into BMSCs efficiently and safely. Adenovirus mediated VEGF(165) gene transferred into BMSCs could express VEGF protein with highly biological activity, which provided foundation on BMSCs based gene therapy for ischemic disease.