Gekle M, Wünsch S, Oberleithner H, Silbernagl S
Department of Physiology, University of Würzburg, Germany.
Pflugers Arch. 1994 Sep;428(2):157-62. doi: 10.1007/BF00374853.
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)
犬肾传代细胞(MDCK)起源于肾集合管,由不同的细胞亚型组成。我们克隆了两种具有不同形态和功能的MDCK细胞亚型,分别命名为C7和C11。这两个克隆在克隆后仍保持其功能差异。C7单层细胞表现出高跨上皮电阻(Rte = 5648 +/- 206 Ω·cm2,n = 20),并向顶端培养基中分泌K+(ΔK+ = 1.31 +/- 0.08 mmol/l,n = 10)。C11单层细胞显示低Rte(330 +/- 52 Ω·cm2,n = 20),并向顶端培养基中分泌Cl-(ΔCl- = 16.9 +/- 1.8 mmol/l,n = 10)。醛固酮(1 μmol/l)刺激C7细胞分泌K+(ΔK+为3.58 +/- 0.11 mmol/l,n = 7),并刺激C11细胞分泌H+(ΔpH = 0.060 +/- 0.007,n = 10)。醛固酮诱导的K+分泌刺激作用可被顶端应用氨氯地平(1 μmol/l)抑制。cAMP刺激C11细胞分泌H+(ΔpH = -0.068 +/- 0.004,n = 10)。此外,C7细胞对花生凝集素(PNA)呈阴性,细胞内pH值为7.39 +/- 0.05(n = 7),而C11细胞将细胞内pH值维持在7.16 +/- 0.05(n = 8),且大部分细胞对PNA呈阳性。我们得出结论,我们克隆了两种稳定表达不同功能特征的MDCK细胞亚型。C7亚型类似于肾集合管的主细胞(PC),而C11亚型类似于肾集合管的闰细胞(ICC)。(摘要截于250字)