Abstract
Objective To detect the expression of VEGF mRNA and protein in rabbit mesenchymal stem cells which stablely transferred by VEGF gene and provide a potential method for treatment of defect and ischemic necrosis of bone.Methods BMSCs were isolated from rabbit bone marrow and cultured in vitro.The vector pcDNA3.1-VEGF165 was transfected into the mesenchymal stem cells by liposome mediated.The stable transferred clone was selected by G418.The mRNA and protein of VEGF165 gene in the transferred cells were analyzed by RT-PCR and Western blot.Results pcDNA3.1-VEGF165 vector was successfully transferred into BMSC and the stable transferred clone was selected.RT-PCR and Western blot showed that there were VEGF165 mRNA and protein expression in the stable transferred cells, but there was no VEGF165 mRNA and protein expression in the other groups.Conclusion pcDNA3.1-VEGF165 vector transfection can obtain the stable expression of exogenous VEGFI65 mRNA and protein in rabbit mesenchymal stem cells, and can be used as the seed cells for the treatment of defect and ischemic necrosis of bone. Key words: Mesenchymal stem cells; Vascular endothelial growth factor; Transfection
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