Transfection of human VEGF165 gene into bone marrow mesenchymal stem cells in rats.

Objective To create a method for transfecting human vascular endothelial growth factor_ 165 (hVEGF_ 165 ) 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 pcDNA_ 3.1 -hVEGF_ 165 was transfected into MSCs with the liposome mediated method. The expression of hVEGF_ 165 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 hVEGF_ 165 in the transfected rat MSCs was demonstrated. Conclusion The vector pcDNA_3.1 -hVEGF_165 is successfully expressed in MSCs.