Characterization of two MDCK-cell subtypes as a model system to study principal cell and intercalated cell properties.
Madin-Darby canine kidney (MDCK) cells originate from the renal collecting duct and consist of different cell subtypes. We cloned two MDCK cell subtypes denominated as C7 and C11 with different morphology and different function. The two clones maintained their functional differences after cloning. C7 monolayers exhibit a high transepithelial resistance (Rte = 5648 +/- 206 omega.cm2, n = 20) and secrete K+ (delta K+ = 1.31 +/- 0.08 mmol/l, n = 10) into the apical medium. C11 monolayers display a low Rte (330 +/- 52 omega.cm2, n = 20) and secrete Cl- (delta Cl- = 16.9 +/- 1.8 mmol/l, n = 10) into the apical medium. Aldosterone (1 mumol/l) stimulates K+ secretion (delta K+ of 3.58 +/- 0.11 mmol/l, n = 7) in C7 cells and H+ secretion in C11 cells (delta pH = 0.060 +/- 0.007, n = 10). Aldosterone-induced stimulation of K+ secretion is inhibited by apical application of amiloride (1 mumol/l). cAMP stimulates H+ secretion in C11 cells (delta pH = -0.068 +/- 0.004, n = 10). Furthermore, C7 cells are peanut-lectin(PNA)-negative and exhibit an intracellular pH of 7.39 +/- 0.05 (n = 7), whereas C11 cells maintain intracellular pH at 7.16 +/- 0.05 (n = 8) and a major fraction of cells is PNA positive. We conclude that we have cloned two subtypes of MDCK cells which stably express different functional characteristics. The C7 subtype resembles principal cells (PC) of the renal collecting duct, whereas the C11 subtype resembles intercalated cells (ICC) of the renal collecting duct.(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Physiology, University of Würzburg, Germany.
M
M
Gekle
S
S
Wünsch
H
H
Oberleithner
S
S
Silbernagl
D000450:Aldosterone / Q000494:pharmacology; D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D002460:Cell Line; D002712:Chlorides / Q000378:metabolism; D002999:Clone Cells; D004285:Dogs; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D017136:Ion Transport / Q000187:drug effects; D007685:Kidney Tubules, Collecting / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D037102:Lectins / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects; D019887:Peanut Agglutinin; D011188:Potassium / Q000378:metabolism; D012964:Sodium / Q000378:metabolism
D002712:Chlorides; D037102:Lectins; D019887:Peanut Agglutinin; D000450:Aldosterone; D003994:Bucladesine; D000584:Amiloride; D012964:Sodium; D011188:Potassium