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.