Comparison of the effectiveness of available sources of autologous colony-forming endothelial cells

Abstract

Endothelial Colony-forming cells (ECFCs) are valuable material for tissue vascular engineering and cell therapy of ischemic tissues. Difficult isolation is the main problem for using of ECFCs. ECFCs isolation from peripheral blood and adipose tissue has been previously described. In the presented research we compared effectiveness of peripheral blood, subcutaneous and epicardial adipose tissue for the ECFCs isolation without cell sorting. ECFCs was isolated from peripheral blood, subcutaneous and epicardial adipose tissue obtained from coronary heart disease patients (males, n=8) undergoing elective coronary artery bypass surgery. The stromal-vascular fraction of subcutaneous (SVF-ST) and epicardial (SVF-ET) adipose tissue as well as the mononuclear blood fraction (MNF) were cultivated in the complied EGM-2 medium. Cell cultures phenotyping was performed by flow cytometry and confocal microscopy. Their angiogenic (Matrigel) and proliferative activity (xCELLigence analyzer) in vitro were studied. ECFCs were isolated from MNF in 50% of cases, from SVF-ST in 12.5% and SVF-ET in 37.5%. The proliferative activity of ECFCs isolated from adipose tissue was low while cultures from MNF showed high and medium activity in 75% of cases. Pure ECFCs (more 99%) were obtained from MNF to third passage without cell sorting. Cultures from adipose tissue were contaminated by mesenchymal-stromal cells (MSCs) and contained ECFCs and MSCs. Thus, peripheral blood is the most effective source of autologous ECFCs compared with adipose tissue for this isolation method. However, adipose tissue is a suitable source of MSC and mixed cultures of MSC and endothelial cells.