The Culture and Differentiation of Endothelial Progenitor Cells from Peripheral Blood in Vitro

Objective To investigate the isolation,culture,differentiation and identification of endothelial progenitor cells(EPCs) from human peripheral blood in vitro.Methods MNCs(mononuclear cells) were isolated from the human peripheral anticoagulant blood sample by Ficoll-density centrifugation,and then the cells were plated on fibronectin-coated culture dishes.The cells were suspended in endothelial basal medium supplemented with ECG-2-MV-SingleQuots.Cultured cells were subjected to immunofluorescence to detect dual binding of Ac-LDL and UEA-1 on day 7.Result Cluster attaching cells appeared in the second day.Cord-like or duct-like structure formed among much more spindle-shaped cells was observed on day 4.EPCs characterized as double positive for Dil-Ac-LDL and FITC-UEI-1.Conclusion EPC can be obtained from mononuclear cells in peripheral blood and be differentiated into endothelial cells at specific conditions in vitro.