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Published on: May 4, 2020
Transfection of human VEGF165 gene into bone marrow mesenchymal stem cells in rats
Jin-fu Yang1, Wen-wu Zhou, Tao Tang
1Department of Cardiothoracic Surgery, Second Xiangya Hospital, Central South University, China. yjf19682005@yahoo.com.cn
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 pcDNA(3.1)-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 pcDNA(3.1)-hVEGF165 is successfully expressed in MSCs.
