ISOLATION AND CHARACTERIZATION OF MACROVASCULAR AND MICROVASCULAR ENDOTHELIAL-CELLS FROM HUMAN HEARTS

In vivo models to investigate mechanisms of local hemostasis in the macro- and microvascular coronary circulation are not available. Therefore, we established a culture system of human macro- and microvascular endothelial cells with high cellular yield and high endothelial cell purity. Microvascular endothelial cells from human hearts were isolated by enzymatic treatment of cardiac muscle preparations obtained during heart transplantation. The isolated microvessels were used to start cultures that were subsequently separated and purified from contaminating nonendothelial cells by paramagnetic beads linked to the lectin Ulex europaeus agglutinin I. Macrovascular endothelial cells were isolated from epicardial coronary arteries and purified by paramagnetic beads as well. With this method high purity (<2% nonendothelial cells) was achieved as judged from fluorescence-activated cell sorting. Immunochemistry demonstrated the expression of several typical endothelial markers. The two endothelial cell, types displayed functional heterogeneity in respect to bradykinin degradation and plasminogen activator inhibitor-1 activity. Thus the ability to selectively isolate and culture human macro- and microvascular cardiac endothelial cells provides a valuable tool to systematically investigate endothelial function in human hearts.