Effects of shear stress on the properties of colonyforming endothelial cells in comparison with coronary artery endothelial cells

Abstract

Highlights. It is assumed that pre-colonization by endothelial cells of the inner surface of tissue-engineered vessels of small diameter can serve as an effective way to prevent thrombosis. The question of choosing the optimal source of endothelial cells for use in tissue engineering remains debatable. The paper considers the features of the culture of colony-forming endothelial cells obtained from the peripheral blood of patients with coronary heart disease, in comparison with mature endothelial cells from the coronary artery.Aim. To study the effect of laminar flow on the morphological and functional characteristics of mature endothelial cells and peripheral blood-derived endothelial colony-forming cells.Methods. Coronary artery endothelial cells were purchased from the Cell Applications, Inc. Colony-forming endothelial cells were obtained from the peripheral blood of patients with coronary artery disease who underwent percutaneous coronary intervention. The cells were isolated using a Ficoll gradient and cultured in EGM-2MV culture medium containing 5% fetal bovine serum. The cells of the experimental group were cultured in µ-Luer plates in a perfusion system with a shear stress of 3 dyn/cm2 . The cultivation time was 2 days. The cells of the control group were cultured under static conditions. At the end of the cultivation we performed immunofluorescent staining for CD31, KDR/CD309, CD144, vWF, type IV collagen, F-actin.Results. Colony-forming endothelial cells and coronary artery endothelial cells retained high density and viability both under static and laminar flow conditions. Shear stress stimulated a change in the phenotype of colony-forming endothelial cells towards a mature endothelial cells, in particular, a significant increased the expression of KDR/CD309 and CD31. The action of laminar flow reduced the synthesis of von Willebrand factor, stimulated the synthesis of type IV collagen. Shear stress promoted the development of structural rearrangements in cells in response to transduction, which manifested in a change in F-actin fibrils orientation on the flow direction.Conclusion. Colony-forming endothelial cells showed a characteristic response to the action of shear stress, consisting in a change in morphology, phenotype, and secretory activity of cells, comparable to that of coronary artery endothelial cells.