Abstract:

BACKGROUND: In recent years, it has been known that endothelial progenitor cells (EPCs) promote the angiogenesis of the body, but the directed differentiation and proliferation of EPCs limits their efficacy and widespread application.
OBJECTIVE: To investigate the feasibility of inducing human peripheral blood derived EPCs into endothelial cells in vitro, and to test their phenotype and function.
METHODS: Mononuclear cells (MNCs) were isolated from the peripheral blood by using Percoll density gradient centrifugation. The cells were then plated on fibronectin-coated culture dishes. After being cultured for 6 days, the attached cells were collected. EPCs were characterized as adherent cells double positive for DiI-LDL-uptake and lectin binding by direct flurescent staining under a laser scanning confocal microscope. The attached cells were cultured and induced by vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in vitro. The morphological changes of the cells were observed. The cells were enumerated by flow cytometry (FCM) with CD31 monoclonal antibody and vWF monoclonal antibody. The attached cells were stimulated with DF-5201 (final concentration: 0.05, 0.1, 0.2 BU/mL) for the respective time points (6, 12, 24 and 48 hours). Proliferation ability of EPCs was assayed by MTT assay.
RESULTS AND CONCLUSION: After induced by VEGF and bFGF in vitro, EPCs became endothelial cell like cells and CD31 and vWF were expressed. Incubation of isolated human MNCs with DF-521 dose- and time-dependently increased the number of EPCs. EPCs can differentiate into endothelial cells by the induction of VEGF and bFGF. Incubation of EPCs with DF-521 increased the proliferative ability at different concentration.