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Am J Physiol Lung Cell Mol Physiol 277: L1096-L1103, 1999;
1040-0605/99 $5.00
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Vol. 277, Issue 6, L1096-L1103, December 1999

Fluid absorption related to ion transport in human airway epithelial spheroids

Peter Steen Pedersen1, Niels-Henrik Holstein-Rathlou2, Per Leganger Larsen3, Klaus Qvortrup4, and Ole Frederiksen2

1 Department of Clinical Genetics and 3 Department of Otolaryngology, Head and Neck Surgery, Rigshospitalet, University of Copenhagen, DK-2100 Copenhagen; and 2 Department of Medical Physiology and 4 Department of Medical Anatomy, The Panum Institute, University of Copenhagen, DK-2200 Copenhagen, Denmark

Airway epithelium explants from cystic fibrosis (CF) patients and non-CF subjects formed monolayered spheres, with the apical ciliated cell membrane facing the bath and the basolateral cell membrane pointing toward a fluid-filled lumen. With the use of two microelectrodes, transepithelial potential difference and changes in potential difference in response to passage of current pulses were recorded, and epithelial resistance and the equivalent short-circuit current were calculated. Non-CF control potential difference and short-circuit current values were significantly lower than the CF values, and amiloride inhibited both values. Fluid transport rates were calculated from repeated measurements of spheroid diameters. The results showed that 1) non-CF and CF spheroids absorbed fluid at identical rates (4.4 µl · cm-2 · h-1), 2) amiloride inhibited fluid absorption to a lower residual level in non-CF than in CF spheroids, 3) Cl--channel inhibitors increased fluid absorption in amiloride-treated non-CF spheroids to a level equal to that of amiloride-treated CF spheroids, 4) hydrochlorothiazide reduced the amiloride-insensitive fluid absorption in both non-CF and CF spheroids, and 5) osmotic water permeabilities were equal in non-CF and CF spheroids (~27 × 10-7 cm · s-1 · atm-1).

amiloride; hydrochlorothiazide; osmotic permeability; current-voltage relationship; cystic fibrosis





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