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.