In vitro study on isolation and induced differentiation of human peripheral blood endothelial progenitor cells

Objective To investigate isolated culture of human peripheral blood vascular endothelial progenitor cells (EPCs) and mononuclear cells (MNCs), and identify that MNCs could be differentiated into EPCs. Methods Human peripheral blood EPCs and MNCs were isolated by Ficoll density gradient centrifugation. MNCs were inducted to differentiate into EPCs with cytokine in vitro. These differentiated EPCs were determined and quantified by immuno-fluorescence and flow cytometer. Results Most of the peripheral blood EPCs and MNCs were proliferated adhesively after being cultured for 3 days. Spindle cells were significantly increased after being cultured for 6 days. After 12 days all cultured cells were harvested for identification of EPCs markers. The uptake rate of Dil-AC-LDL was (70.00±11.21)% in cultured cells, and the cytoplasm in most of the attaching cells were positive for AC133, CD31, CD34, vascular endothelial growth factor receptor-2 (Flk-1) and von Willebrand factor (vWF) in immuno-fluorescence. Flow cytometric analysis showed that the positive rates of AC133, CD31, CD34 and Flk-1 of attaching cells were 95.80%, 87.60%, 1.27% and 96.40% respectively. Conclusion This study suggests that EPCs differentiated from MNCs by induction have the characteristics of progenitor cells and the induction rate is high when they are cocultured in vitro.