Primary culture of collecting duct cell epithelium from neonate rabbit kidney in monolayer

Abstract

SUMMARYCortical 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 ,-cell-probe, peanut agglutinin. Single cells were isolated by brief exposure to 0 Ca2l-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 /LM-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.

Flow cytometric immunodissection of the human distal tubule and cortical collecting duct system

“…enriched segments in rabbits [20,21], but not humans. Several groups have used the microdissection approach Nouwen and De Broe demonstrated that EGF immunostaining using polyclonal AB-3 results in distinct cellular to isolate and purify individual subsegments leading to pure cultures of TAL cells, DCT cells and/or CT/CD staining patterns in different (sub)segments [TAL, predominant apical surface staining; DCT, cytoplasmic cells in rats, rabbits [5, [22][23][24][25], and humans [6, 26,27]. Manual microdissection allows high-grade purification, staining; early and late CT, staining equally distributed over the entire cell surface on Ͼ75% and Ͻ50% of cells but the cell yield is relatively low.…”