Abstract:

BACKGROUND: Traditional method for culture of endothelial progenitor cells from human peripheral blood is complex to operate, costs high, and acquires a small number of cells.
OBJECTIVE: To culture endothelial progenitor cells from human peripheral blood using autologous serum and identify their functions.
METHODS: Peripheral blood was collected from volunteers. Mononuclear cells were separated by density centrifugation and were induced to differentiate into endothelial progenitor cells in vitro. The influences of different culture conditions (EGM-2MV, M199+10% autologous serum+insulin like growth factor, M199+10% fetal bovine serum+vascular endothelial growth factor+basic fibroblast growth factor+insulin-like growth factor+epidermal growth factor) on the proliferation and migration of endothelial progenitor cells were observed. The cultured endothelial progenitor cells were identified by cell morphology observation, fluorescent staining and flow cytometry.
RESULTS AND CONCLUSION: Endothelial progenitor cells with higher proliferation and migration ability were obtained under EGM-2MV and M199+10% autologous serum + insulin-like growth factor conditions (P < 0.05). Furthermore, the percentage of double positive cells stained by Dil-labeled acetylated low density lipoprotein and FITC-labeled Ulex europaeus agglutinin 1 was higher than 80%, and these cells also expressed CD34, CD133 and KDR. Addition of autologous serum to M199 culture medium is a simple, highly efficient method for culturing endothelial progenitor cells.