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Primary culture of collecting duct cell epithelium from neonate rabbit kidney in monolayer
Author(s) -
White S,
Reeve H
Publication year - 1992
Publication title -
experimental physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.925
H-Index - 101
eISSN - 1469-445X
pISSN - 0958-0670
DOI - 10.1113/expphysiol.1992.sp003567
Subject(s) - amiloride , explant culture , intercalated cell , cell culture , epithelium , biology , duct (anatomy) , microbiology and biotechnology , cell , kidney , chemistry , anatomy , endocrinology , biochemistry , in vitro , sodium , genetics , organic chemistry
Cortical collecting duct cells from rabbit neonate kidney capsula fibrosa explants were grown in low (0.1%) serum medium on permeable culture supports. We found that over a period of 3–4 days in culture, the cells migrated from the explants, to form monolayers. The cells in monolayer displayed morphology similar to adult collecting duct cells. Approximately 24% of cells grown in monolayer were found to bind a specific beta‐cell‐probe, peanut agglutinin. Single cells were isolated by brief exposure to 0 Ca(2+)−Mg2+ solutions and the whole‐cell clamp technique was used to measure macroscopic currents. The whole‐cell conductance of 1.35 +/− 0.1 nS was reduced to 0.9 +/− 0.2 nS in the presence of 100 microM‐amiloride. We conclude that these cells express cell membrane properties consistent with those of the collecting duct and that it may be a useful model for the study of cellular transport mechanisms and the factors controlling development of collecting duct cell types.