[Expression of human VEGF165 gene in rat bone marrow stromal cells in vitro].
OBJECTIVE
The purpose of this study was to evaluate the feasibility of VEGF165 gene transfection into bone marrow stromal cells and make the groundwork of VEGF gene therapy for the revascularization of bone tissue engineering.


METHODS
Bone marrow stromal cells (BMSC) were cultured in vitro, and were tested for the dexamethasone-induced potential to form mineralized-like tissue in culture in the presence of ascorbic acid and beta-glycerophosphate. The vector pcDNA3.1-VEGF165 was transfected into bone marrow stroma cells with the method of liposome mediated or Sofast transfectam reagent. The expression level of human VEGF165 in the transformed cells was detected by immunocytochemistry.


RESULTS
The percent of the positive cells after VEGF165 gene transfection mediated by liposome and cationic polymer was 11.34 and 11.42 respectively. There was no significant difference in gene transfer efficacy between liposome and cationic polymer mediated gene delivery. While the percent of the number of survival cells after gene transfection was 74.60 and 85.88 respectively, the cytoxity of liposome was a little higher than that of cationic polymer.


CONCLUSIONS
The experiment demonstrated that rhVEGF165 was successfully expressed in BMSC. This result could serve as a basis for the next step of revascularization of bone tissue engineering.
Bai
Xiao-feng
XF
Department of Oral and Mazillofacial Sugery, West China School of Stomatology, Sichuan University, Chengdu 610041, China.
Tian
Wei-dong
WD
Chen
Xi-zhe
XZ
Li
Zhi-yong
ZY
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D008081:Liposomes; D051381:Rats; D017207:Rats, Sprague-Dawley; D017154:Stromal Cells / Q000166:cytology* / Q000378:metabolism; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis / Q000235:genetics*
D008081:Liposomes; D042461:Vascular Endothelial Growth Factor A