Characterization of the glomerular endothelial cell in culture

Because of difficulties associated with the culture, cloning and propagation of glomerular endothelial cells (GENs), the biological properties of these cells remain largely unknown. We modified the methods established by Ballermann to propagate GENs from adult bovine kidney. We found that the addition of insulin, transferrin and selenium into the standard culture media was an important step in promoting the propagation of the first clone from a single cell and in maintaining the viability of the cells. These cells expressed factor VIII-related antigen and took up acetylated-LDL, but did not contain the Weibel-Palade body, unlike endothelial cells derived from large vessels. Furthermore, GENs were compared with aortic endothelial cells (AECs) to investigate the differences in culture conditions. Compared with AECs, GENs required a higher concentration of serum and the supplementation of growth factor to maintain their biological activity. In addition, GENs were very susceptible to trypsinization and produced prostaglandin E2 as a major cyclooxygenase product, whereas AECs produced PGI2. These findings suggest that GENs will be easily obtained from adult bovine kidney in culture and provide useful information on the functional properties of these cells under physiological and pathophysiological conditions.