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Na+ transport activity was characterized in human cheek epithelial cells obtained from normotensive adult subjects. The cells were isolated using a mouth-wash procedure and assayed for Na+ uptake using a radioactive (22Na+) rapid filtration assay. Cheek cells displayed proton-dependent Na+ uptake activity that was dependent on the magnitude of the externally directed proton gradient measured using the fluorescent probe 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein to determine intracellular pH. Amiloride, ethylisopropylamiloride (EIPA), 5-(N,N-dimethyl)-amiloride, 5-(N-methyl-N-isobutyl)-amiloride (MIA), and 5-(N,N-hexamethylene)-amiloride (NNHA) all inhibited proton-dependent Na+ uptake, with MIA, EIPA, and NNHA being the most potent. The Michaelis constant (Km) for extracellular Na+ was 5.7 mM, while the maximum velocity for Na(+)-H+ antiporter activity was 4.3 nmol Na+.mg protein-1.30s-1. The Km for intracellular H+ was 0.17 microM, with a Hill coefficient of 0.7. Stimulation by ouabain and inhibition by bumetanide of cheek cell proton-dependent Na+ uptake indicated only relatively low activities of Na(+)-K(+)-ATPase and Na(+)-K(+)-2Cl- cotransport, respectively. These results are consistent with the presence of Na(+)-H+ antiporter activity in cheek cells. Cheek cells therefore provide a convenient, relatively noninvasive source of tissue for examining Na(+)-H+ antiporter activity in human subjects.

This study describes some biological properties of human cheek (buccal epithelial) cells, isolated by mouth wash. Yields ranged from 6.5 to 20.6 x 10(6) cells, with a mean (+/- SE) of 12.2 +/- 4.2 x 10(6) cells, which gave 0.55 +/- 0.01 x 10(6) cells/mg protein. Vital stain exclusion was similar in cells isolated in either water (89 +/- 2%) or 250 mM sucrose (87 +/- 3%). From our measurements of cell volume and electrolyte content, we estimated intracellular Na+ and K+ concentrations to be between 0.3-0.5 and 7.4-13.0 mM, respectively. In 22 adult subjects, basal, prickle, intermediate, and superficial cells represented 0.3 +/- 1.4, 51 +/- 2.4, 26 +/- 0.9, and 22.7 +/- 1.8%, respectively, of the total sample. Cheek cells exhibited a low endogenous rate of oxygen consumption, which was stimulated by glucose or succinate and inhibited by KCN or NaF. Cheek cells were osmotically stable in a wide range of media, including water. However, they exhibited shrinkage and collapse in hypertonic media, particularly polyethylene glycol.