Characterization and culture of sinusoidal endothelium from normal rat liver: lipoprotein uptake and collagen phenotype

Abstract

Sinusoidal endothelial cells from adult rat liver have been isolated by centrifugal elutriation and been established in primary culture. Their identification has made use of a novel process of these cells, the sequestration of human acetoacetylated low-density lipoprotein. After its administration in vivo, labeled acetoacetylated low-density lipoprotein within the liver was associated solely with sinusoidal endothelial cells and, when labeled with the stable fluorescent compound 3,3'-dioctadecylindocarbocyanine, provided a means of identifying isolated cells by fluorescence microscopy. In endothelial cell cultures, 90% of the cells were fluorescent and exhibited fenestrae. The collagen phenotype of cultures was assessed by an immunofluorescent approach, which revealed cell-associated type IV collagen only; types I, III, and V were undetectable. In other studies, it was found that these cells lacked factor VIII-R antigen and Weibel-Palade bodies, adding to the evidence that they differ substantially from large-vessel and other capillary endothelia.