Abstract:AimTo investigate the effects of Jagged1 over-expression on aged rat-derived endothelial progenitor cells migration and proliferation.MethodsMononuclear cells were obtained from bone marrow of young (1 to 2 month-old) and aged (19 to 26 month-old) Sprague-Dawley rats by Ficoll density gradient centrifugation and cultured with medium DMEM/F12, 48 h later, the suspending cells were translocated to be cultured in new flasks coated with fibronectin, the secondary attached cells were used to perform the further experiments.EPCs were characterized as double positive for Dil-ac-LDL uptake and lectin binding and cells were further identified by vWF expression.The experiments were divided into four groups: control group (aged rat-derived EPCs without gene transfection), PIRES2-EGFP transfection group, PIRES2-EGFP-Jagged1 transfection group and young rat-derived EPCs group in which transfection was also not performed.The GFP expression cell number was acquired through fluorescence microscope and transfection efficiency was calculated.Immunofluorescence, RT-PCR and Western blot were used to detect the mRNA and protein expression respectively.Cell migration and proliferation were tested by Transwell culture and MTT assay respectively.ResultsImmunofluorescence showed that gene transfection markedly promoted Jagged1 expression; RT-PCR and Western blot demonstrated that Jagged1 mRNA and protein expression significantly increased in aged rat-derived EPCs (P<0.01) compared