An osmotically tolerant inner medullary collecting duct cell line from an SV40 transgenic mouse

The terminal inner medullary collecting duct (IMCD) plays an important role in determining the final urinary composition. Currently, there is no continuous cell line derived from this nephron segment. We have developed a cell line derived from the terminal IMCD of mice transgenic for the early region of simian virus SV40 (large T antigen). This cell line, mIMCD-3, retains many differentiated characteristics of this nephron segment including high transepithelial resistance (1,368 ± 172 Ω · cm²), inhibition of apical-to-basal sodium flux by amiloride (41 ± 7%) and by atrial natriuretic peptide (ANP) (40 ± 9%), the presence of the amiloride-sensitive sodium channel as determined by Western blot analysis, and accumulation of the major organic osmolytes in response to hypertonic stress. Significantly, mIMCD-3 cells adapted readily and were able to grow in hypertonic medium supplemented with NaCl and urea up to 910 mosmol/kgH₂O. These extreme osmotic conditions exist in the renal medulla in vivo but are known to be lethal to most other cells. This cell line should be highly useful for the study of the cellular adaptation to osmotic stress and the cell biology and transport physiology of this nephron segment. hyperosmotic stress; organic osmolytes; sodium channel; atrial natriuretic peptide; renal medulla; dexamethasone; ZO-1; desmoplakin