Expression of vascular endothelial growth factor-165 in endothelial progenitor cells transfected with pcDNA3.1(+)/VEGF165 plasmid / 重庆医科大学学报

ABSTRACT

Objective:To culture and identify ratendo the lialprogen it or cells,and too bserve the expression of recombin anthuman vascular endothelial growth fator-165(VEGF165)in endothelial progenitor cells transfected with pcDNA3.1(+)/VEGF165 plasmid.Methods:Endothelial progenitor cells were cultured in vitro,and immunocytochemical staining was used to detect the expression of surface antigen(CD34,CD133,KDR).After transfection of VEGF165,VEGF protein in the culture supernatant of endothelial progenitor cells was detected by enzyme-linked immunosorbent assay(ELISA),and VEGF165 mRNA in endothelial progenitor cells was detected by reverse transcription polymerase chain reaction(RT-PCR).Results:The cultured cells were identified to be endothelial progenitor cells by immunocytochemical analysis.Gel electrophoresis showed that a 576 bp product was amplified by RT-PCR in VEGF165-transfected cells,while a weak expression of VEGF165 was observed in mock-transfected and non-transfected cells.The protein level of VEGF165 in the supernatant of VEGF165-transfected,mock-transfected and non-transfected cells were 175.8?10.7 pg/ml,10.5?1.6 pg/ml and 9.3?1.3 pg/ml,respectively.Conclusion:There is small amount of VEGF165 expression in endothelial progenitor cells in vitro,and the endothelial progenitor cells transfected with pcDNA3.1(+)/VEGF165 plasmid have an increased expression of VEGF165.