An in vitro study of PcDNA3.1 (+)NEGF165 transfection on endothelial progenitor cell derived from murine bone marrow / 中华内分泌外科杂志

ABSTRACT

Objective To investigate the effect ofpcDNA3.1 (+)/VEGF165 plasmid transfection on endothelial progenitor cells (EPCs)derived from murine bone marrow mediated by liposome.Methods EPCs were isolated from murine bone marrow cuhured in vitro,identified by immunocytochemistry and fluorescence-activated cell sorting.After transfection,VEGF165 mRNA in EPCs was detected by reverse transcription polymerase chain reaction (RT-PCR),and VEGF protein in the culture supernatant of EPCs was detected by enzyme-linked immunosorbent assay (ELISA).MTT method was used to study the effect of transfection by pcDNA3.1 (+)/VEGF165 plasmid on EPCs.Results EPCs expressed cell markers CD34,CD133,KDR.Gel electrophoresis showed that a 576bp 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 was higher than that in the other groups.VEGF165 plasmid transfer could stimulate EPCs proliferation.Conclusion EPCs can successfully transfect VEGF165 gene and promote the proliferation of EPCs,providing experimental basis for treatment of erectile dysfunction.