CDC/EU.HMEC-1 (HMEC-1) cells provide a reliable source of human micro-vascular endothelial cells free of mycoplasma and viral infection. This cell line has potential for use in the further study of the endothelial cell modification of low-density lipoproteins and for anticholesterol drug evaluation assays. HMEC-1 cells will fill a gap that is present for in vitro investigations of cholesterol metabolism in conjunction with previously established hepatic, monocytic, or macrophage cell lines. This paper presents a simple assay that demonstrates a linear uptake of tritiated cholesterol by he HMEC-1 cells and shows that the cellular cholesterol load can be regulated using anticholesterol drugs.