Isolation of metabolically active endothelial cells in high yield from bovine cavernous bodies. A model for functional studies on freshly isolated microvascular endothelial cells

Abstract

Work on endothelial cells has been limited by the availability of procedures for obtaining such cells in quantities adequate for direct in vitro analysis. The present paper describes a method for the isolation of endothelial cells from bovine cavernous bodies. A number of cells ranging from 2.5 to 4 X 10(8) per animal has been obtained. The cells were identified as follows 1) presence of the "Weibel and Palade" bodies in the isolated cells, 2) "cobblestone" appearance of cell cultures, and 3) presence of factor VIII, as demonstrated by immunofluorescence assays. The cell viability at the end of the purification procedure was tested 1) by dye-exclusion tests and 2) by metabolic assays. Features of this preparation are 1) the very high yield of viable endothelial cells, 2) the absence of contamination by fibroblasts and smooth muscle cells and a very low contamination by erythrocytes and 3) the fine dispersion of the isolated cells. These properties allow functional and subcellular fractionation studies on freshly isolated endothelial cells of microvascular origin.