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Am J Physiol Lung Cell Mol Physiol 274: L450-L453, 1998;
1040-0605/98 $5.00
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Vol. 274, Issue 3, L450-L453, March 1998

Evidence that Calu-3 human airway cells secrete bicarbonate

Michael C. Lee1, Christopher M. Penland1, Jonathan H. Widdicombe2,3, and Jeffrey J. Wine1

1 Cystic Fibrosis Research Laboratory, Stanford University, Stanford 94305-2130; 2 Cardiovascular Research Institute, University of California, San Francisco 94143; and 3 Children's Hospital Oakland Research Institute, Oakland, California 94609

The Calu-3 cell line is being investigated as a model for human submucosal gland serous cells. In a previous investigation of basal short-circuit current (Isc) in Calu-3 cells, high levels of bumetanide-insensitive basal Isc (~60 µA/cm2) were measured in cells grown at an air interface. Basal Isc was reduced only 7% by bumetanide, and the largest component of basal Isc required both Cl- and HCO<SUP>−</SUP><SUB>3</SUB> in the bathing solutions. Because Isc could be partially inhibited by basolateral 4,4'-dinitrostilbene-2,2'-disulfonic acid and because the only known apical exit pathway for anions is the cystic fibrosis transmembrane conductance regulator, which has a relatively poor conductance for HCO<SUP>−</SUP><SUB>3</SUB>, it was concluded that most basal Isc is HCO<SUP>−</SUP><SUB>3</SUB>-dependent Cl- secretion [M. Singh, M. Krouse, S. Moon, and J. J. Wine. Am. J. Physiol. 272 (Lung Cell. Mol. Physiol. 16): L690-L698, 1997]. We have now measured isotopic fluxes of 36Cl- and 22Na+ across short-circuited Calu-3 cells and found that virtually none of the basal Isc is Cl- secretion or Na+ absorption. Thus, in contrast to the earlier report, we conclude that the major component of basal Isc is HCO<SUP>−</SUP><SUB>3</SUB> secretion. Stimulation recruits primarily Cl- secretion, as previously proposed.

cystic fibrosis; Ussing chamber; epithelia; submucosal gland; cell culture


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