Visceral glomerular epithelial cells (v-GECs) are highly differentiated and specialized cells which form an important component of the glomerular filtration barrier. As v-GECs are highly differentiated cells, it is not elucidated whether v-GECs have the capacity to proliferate and whether GEC grown in culture are of visceral or parietal origin. We found this difference to be related to difficulty in culturing GEC beyond the 6th passage. With immunocytochemical methods, the cells which were difficult to culture were found to possess characteristics of v-GECs. In these cells, we investigated the mechanisms of cell death, particularly the appearance of apoptosis. Under serum free conditions, v-GECs degenerated in a time dependent manner. With flow cytometric analysis and agarose gel electrophoresis, cell death under serum free conditions and after 6th passages depended on apoptotic mechanisms based on the demonstration of DNA fragmentation. The addition of heparin and protein kinase C inhibitors, H-7 and staurosporin, suppressed cell degeneration and DNA fragmentation. It was concluded that v-GECs could not be cultured beyond the 6th passage due to apoptotic mechanisms and the development of apoptosis appeared to require the activation of protein kinase C
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