Effects of rapamycin on number and activity of endothelial progenitor cells / 解放军医学杂志

ABSTRACT

Objective To investigate the effects of rapamycin on the number and function of peripheral blood 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-day culture, adherent cells were treated with rapamycin in a series of final concentrations of 1.0, 2.0, 5.0?g/ml for 6, 12, 24, and 48h. EPCs were identified as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining as demonstrated under a laser scanning confocal microscope. EPCs were further documented by demonstrating the expression of VEGFR-2, AC133 and CD34 with flow cytometry. EPCs proliferation and migration were assayed with MTT assay and modified Boyden chamber assay, respectively. EPCs adhesion assay was performed by replating them on fibronectin-coated dishes, and then adherent cells were counted. In vitro vasculogenesis activity was assayed by in vitro vasculogenesis kit. Results Incubation of isolated human MNCs with rapamycin resulted in a decrease in the number of EPCs, and rapamycin also decreased EPCs proliferative, migratory, adhesive and in vitro vasculogenesis capacity in both concentration and time dependent manners. Conclusion Rapamycin decreases the number, proliferative, migratory, adhesive and in vitro vasculogenesis capacity of EPCs.