Culture of human nasal epithelial cells on collagen matrix supports. A comparison of bioelectric properties of normal and cystic fibrosis epithelia.
The feasibility of culturing human nasal epithelial cells was tested by plating on permeable collagen matrices primary isolates of epithelial cells from normal (n = 21) and atopic (n = 5) subjects and from subjects with cystic fibrosis (CF) (n = 13). The cells were cultured in serum-free, hormone-supplemented medium for 5 days followed by maintenance in conditioned medium with 1% serum. A transepithelial electric potential difference (PD) exceeding 1.0 mV developed in most preparations (71%). The PD reached a peak by Day 10 and declined slowly thereafter. The maximal PD of CF epithelia (-32.4 +/- 5.8 mV, n = 9) on collagen matrices exceeded that of normal (-11.3 +/- 1.8 mV, n = 15) and atopic (-13.5 +/- 2.5, n = 5) epithelia. The maximal PD for both CF and normal cultures exceeded that of freshly excised preparations. This difference reflected higher transepithelial resistance of cultures as compared with intact preparations. The efficacy of amiloride (10(-4) M) to reduce the PD was greater in cultures of CF epithelia (-85 +/- 4.6%, n = 7) than it was in cultures of normal (-45.5 +/- 5.0%, n = 11) and atopic (-50.5 +/- 2.9%, n = 5) epithelia, whereas the hyperpolarization induced by replacement of Cl- in the apical bath was smaller for CF cultures (5.9 +/- 1.5 mV) than it was for normal (9.3 +/- 0.9 mV) and atopic (8.8 +/- 0.5 mV) cultures. We conclude that nasal epithelial cells will form high resistance barriers when cultured on collagen matrices.(ABSTRACT TRUNCATED AT 250 WORDS)
D000293:Adolescent; D000328:Adult; D000368:Aged; D002478:Cells, Cultured; D002648:Child; D003094:Collagen; D003470:Culture Media / Q000494:pharmacology; D003550:Cystic Fibrosis / Q000473:pathology / Q000503:physiopathology; D005260:Female; D006801:Humans; D008297:Male; D008564:Membrane Potentials; D008722:Methods; D008875:Middle Aged; D009297:Nasal Mucosa / Q000502:physiology / Q000648:ultrastructure*; D009298:Nasal Polyps / Q000648:ultrastructure; D006255:Rhinitis, Allergic, Seasonal / Q000473:pathology / Q000503:physiopathology
D003470:Culture Media; D003094:Collagen