Culture and differentiation of endothelial progenitor cells from peripheral blood in vitro

Objective To investigate the methods of isolating, culturing, differentiating and evaluating the endothelial progenitor cell (EPC ) . Methods The mononuclear cells were obtained by Ficoll density-gradient centrifu-gation. The cells were suspended in endothelial basal medium (EBM-2) supplemented with EGM-2-MV-SingleQuots for culturing, differentiating and proliferating. The expressions of specific antigens on cell surface were analysed by immunohistochemistry and immunofluorescence. Results The EPCs exhibited the clonal morphology and classical cobblestone morphology after 10 and 20 days culture respectively. The endothelial phenotype was confirmed by positive immunostaining with endothelial cell-specific markers, such as CD31, vWF, FLK-1/KDR/VEGFR-2 , and VE-cadherin. Conclusion EPC can be obtained from mononuclear cells in peripheral blood and be differentiated into endothelial cell in vitro.