Ion transport by primary cultures of canine tracheal epithelium: methodology, morphology, and electrophysiology.
Canine tracheal epithelial cells were isolated by enzymatic and mechanical dispersion and cultured on permeable supports. The cells formed confluent monolayers and retained most of the morphologic characteristics of the intact epithelium, including apical microvilli, apical tight junctions, and a moderately interdigitated lateral intercellular space. The cells also retained the functional properties of the epithelium. The monolayer responded to addition of isoproterenol with the characteristic changes in cellular electrical properties expected for stimulation of C1 secretion: isoproterenol increased transepithelial voltage, depolarized apical membrane voltage, and decreased both transepithelial resistance and the ratio of apical-to-basolateral membrane resistance. Examination of the cellular response to ion substitutions and inhibitors of C1 secretion indicate that the cultured monolayers retain the same cellular mechanisms of ion transport as the intact epithelium. Thus, primary cultures of tracheal epithelium may provide a useful preparation for future studies of the mechanism and regulation of C1 secretion by airway epithelia.
D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects; D002462:Cell Membrane / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D002463:Cell Membrane Permeability / Q000187:drug effects; D002469:Cell Separation; D002478:Cells, Cultured; D002712:Chlorides / Q000378:metabolism*; D004285:Dogs; D004553:Electric Conductivity; D004848:Epithelium / Q000378:metabolism / Q000648:ultrastructure; D005614:Freeze Fracturing; D007545:Isoproterenol / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects; D008854:Microscopy, Electron; D010042:Ouabain / Q000494:pharmacology; D013997:Time Factors; D014132:Trachea / Q000378:metabolism* / Q000648:ultrastructure
D002712:Chlorides; D010042:Ouabain; D007545:Isoproterenol