Isolation and Culture of Human Peripheral Blood Mononuclear Cells for Differentiating into Endothelial Progenitor Cells and Endothelial Cells

Objective To investigate the methods of isolation and culture of human peripheral blood mononuclear cells(PBMC)for differentiating into endothelial progenitor cells(EPCs)and endothelial cell(ECs).Method Peripheral blood mononuclear cells were isolated by density-gradient centrifugation and incubated in endothelial medium in the presence of vascular endothelial growth factor.Peripheral blood mononuclear cells were differentiated into endothelial progenitor cells and endothelial cell.Flow cytometry and PT-PCR were used to identify the endothelial progenitor cells and endothelial cell.Results After 7 days,most of the cultured cells displayed a fibroblast-like morphology adhering to the culture plate.They expressed CD34,CD133 and VEGFR.After 4 weeks,most of the cultured cells expressed vWF and eNOS.Conclusion The endothelial progenitor cells and endothelial cell were successfully cultured from peripheral blood mononuclear cells