Mechanical Isolation of Human Endothelial Cells from Large Vessels for Flow Cytometry Immunophenotyping

Abstract

Endothelial cells (EC) have important physiological functions, and they may also have a role in pathology. To better understand their role in health and disease, we must know very well their phenotype. Previous studies have identified and characterized EC mainly by immunohistochemistry, but there are also some studies using flow cytometry (FCM) after exposing these cells to enzymatic digestion, either to isolate and/ or to detach them from the vessel wall. However, it is well known that enzymatic treatment can cause deleterious effects on cell surface receptors, then influencing the antibody-antigen reaction. We describe a simple and cheap mechanical method to isolate EC from human vessels, avoiding alterations in the expression of cell surface receptors caused by the use of enzymes, and we tested it using FCM. With this method we were able to obtain fresh EC that were identified by FCM as a well-defined cluster of CD45 - CD146+bright CD31 +bright cells. This approach can be used in the future to isolate EC for further immunophenotypic characterization and for ex-vivo functional studies, as well as to test the effect of different stimuli, including pharmacological drugs.

Morphological restructuring of smooth muscles elements of the wallgreat saphenous vein of a person on the lower leg during the development of varicose transformation

“…Большинство исследований в отношении развития эндотелиальной дисфункции на сегодня посвящено изучению взаимного влияния ультраструктурных изменений, наступаю щих в эндотелиальных клетках, на количественную и качественную характеристику выделяющихся в кровь эндогенных эндотелиальных факторов, влияющих на проницаемость сосудистой стенки и развитие воспалительных реакций или активизацию факторов тромбoобразования [12,32]. В связи с чем необходимо отметить, что если роль эндогенных эндотелиальных факторов в развитии тромбоза вен на сегодня установлена почти полностью, то в отношении непосредственного развития варикозной болезни дискуссия пока далека от своего завершения [17,[33][34][35][36][37].…”