Transfection of human VEGF165 gene into bone marrow mesenchymal stem cells in rats / 中南大学学报(医学版)

ABSTRACT

Objective To create a method for transfecting human vascular endothelial growth factor165 (hVEGF165) gene into bone marrow mesenchymal stem cells (MSCs) in rats. Methods MSCs of Wistar rats were isolated by density gradient centrifugation and purified based on their ability of adhesion to plastic. Detections of cell surface antigens, including CD34, CD45, CD44, and SH3, were performed using flow cytometry. MSCs' potential of differentiating into osteoblast and lipoblast in vitro was tested. The vector pcDNA3.1-hVEGF165 was transfected into MSCs with the liposome mediated method. The expression of hVEGF165 in the transfected cells was detected by enzyme linked immunosorbent assay (ELISA), reverse transcription-polymerase chain reaction (RT-PCR), and Western blot analysis. Results The cultured MSCs were CD34-, CD45-, CD44+, and SH+, which were differentiated into osteoblasts and lipocytes successfully. The expressed hVEGF165 in the transfected rat MSCs was demonstrated. Conclusion The vector pcDNA3.1-hVEGF165 is successfully expressed in MSCs.