title: Transfection of human VEGF165 gene into bone marrow mesenchymal stem cells in rats.
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 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.
authors.affiliation: Department of Cardiothoracic Surgery, Second Xiangya Hospital, Central South University, China. yjf19682005@yahoo.com.cn
authors.forename: Jin-fu
authors.initials: JF
authors.lastname: Yang
authors.forename: Wen-wu
authors.initials: WW
authors.lastname: Zhou
authors.forename: Tao
authors.initials: T
authors.lastname: Tang
authors.forename: Jian-guo
authors.initials: JG
authors.lastname: Hu
authors.forename: Jie-feng
authors.initials: JF
authors.lastname: Yu
authors.forename: Yi-feng
authors.initials: YF
authors.lastname: Yang
authors.forename: Xin-min
authors.initials: XM
authors.lastname: Zhou
authors.forename: Dong-xu
authors.initials: DX
authors.lastname: Hu
mesh_terms: 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
chemical_list: D018952:Antigens, CD34; D018960:Hyaluronan Receptors; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D017493:Leukocyte Common Antigens