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


CONCLUSIONS
The vector pcDNA(3.1)-hVEGF165 is successfully expressed in MSCs.
Yang
Jin-fu
JF
Department of Cardiothoracic Surgery, Second Xiangya Hospital, Central South University, China. yjf19682005@yahoo.com.cn
Zhou
Wen-wu
WW
Tang
Tao
T
Hu
Jian-guo
JG
Yu
Jie-feng
JF
Yang
Yi-feng
YF
Zhou
Xin-min
XM
Hu
Dong-xu
DX
D000818:Animals; D018952:Antigens, CD34 / Q000032:analysis; D001854:Bone Marrow Cells / Q000166:cytology; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D006801:Humans; D018960:Hyaluronan Receptors / Q000032:analysis; D017493:Leukocyte Common Antigens / Q000032:analysis; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis* / Q000235:genetics
D018952:Antigens, CD34; D018960:Hyaluronan Receptors; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D017493:Leukocyte Common Antigens