Abstract
Endothelial cells are a major contributor to cardiovascular diseases. Studying their function and how they influence pathological processes remains an ongoing area of research. Although primary endothelial cells might be readily available from animals, translating results from these studies can be challenging due to species-dependent differences. A common source of human endothelial cells is human umbilical vein endothelial cells, but these cells might not represent the variety of endothelial cells from various vascular beds. Here we describe a protocol for the isolation and cultivation of endothelial cells from human internal mammary artery remains. These remains are commonly available from coronary artery bypass graft surgeries. The described tissue explant method combined with a magnetic sorting process provided a relatively high yield of endothelial cells that were subsequently passaged and used for studies involving mRNA and protein expression analyses, as well as fluorescence-based immunohistochemistry and patch-clamp electrophysiology. This protocol may help to extend the repertoire of studying human endothelial cells to understand their role and function in health and disease.