Effects of high glucose on number and activity of endothelial progenitor cells from peripheral blood / 解放军医学杂志

ABSTRACT

Objective To investigate the effect of high glucose on the number and proliferation, migration and adhesion of peripheral endothelial progenitor cells (EPCs). Methods Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin coated culture dishes. After 7 days of culture, several groups of attached cells were incubated with glucose in a series of concentrations (15, 25, 35, 45mmol/L) for different durations (6, 12, 24 and 48h). EPCs were characterized as adherent cells which were double positive for DiLDL uptake and lectin binding by direct fluorescent staining demonstrated under a laser scanning confocal microscope. EPCs were further documented by demonstrating the expression of KDR, VEGFR 2 and AC133 with flow cytometry. EPCs proliferation, migration and in vitro vasculogenesis activity were assayed with MTT assay, modified Boyden chamber assay and in vitro vasculogenesis kit, respectively. EPCs adhesion assay was performed by replating MNCs on fibronectin coated dishes, and then the adherent cells were counted. Results Incubation of isolated human MNCs with high glucose concentration decreased the number of EPCs, and this effect was most prominant when glucose concentration was 45mmol/L, and incubated for 24 hours (approximately 1 fold decrease, P