Korean Journal of Nephrology 1995;14(4):433-446.

백서의 사구체 격측상피세포의 배양가 성상확인

하태선 , 최용

Abstract

For the purpose of evaluating the origin of cultured glomerular epithelial cells(GEC), we applied immunohistochemical(IHC) and electron microscopic (EM) studies to characterize the cultured rat GEC. After isolation by differential sieving method, glomeruli were explanted and maintained in a medium(K1-3T3) consisting of equal parts of a Kl medium and conditioned medium taken from cultures of NIH 3T3 fibroblasts grown in Dulbecco's modified Eagle's mediurn plus 10% fetal calf serum. Morphologically distinct GEC grew out from the isolated glomeruli and they were cultured continuously. The grown GEC(early stage within 7 days after glomerular explantation) and continuously cultured cells were characterized by IHC using primary antibodies, such as, monoclonal antibodies to cytokeratin, desmin, Thy 1.1, and vimentin and EM methods. At the sarne time, their results were compared with those of glomerular epithelium from rat kidney sections. The inverted light microscopic features of continuously cultured GEC were similar to those of primarily cultured GEC. Both prirnarily and continuously cultured GEC stained positive for cytokeratin and negative for desmin, Thy 1.1, and vimentin, which results were identical to the IHC results of parietal cells in situ. On the contrary, visceral epithelial cells in situ stained positive for vimentin and desmin and negative for cytokeratin. These results suggested that rat GEC in primary and continuous culture, expressed IHC characteristics of in situ parietal, not visceral epithelium. We could observe junctional complexes of the desmosome type in cultured GEC which were noted in parietal epithelial cells, not in visceral cells. In conclusion, the cultured epithelial cells might be glomerular parietal epithelial cells by comparing the IHC and EM characteristics of cultured GEC with glomerular epithelium from rat kidney sections. Therefore, we can use these cultured and characterized glomerular parietal epithelial cells (GPEC) for the study of their biologic functions or of the pathophysiology of GPEC-associated diseases, such as, crescentic glomerulonephritis.