Electrical properties of monolayers cultured from cells of human tracheal mucosa

Jonathan Widdicombe, D. L. Coleman, W. E. Finkbeiner, I. K. Tuet

Research output: Contribution to journalArticle

45 Scopus citations

Abstract

Dispersed isolated cells were obtained from human tracheal mucosa by digestion with collagenase. Up to 1.5 x 108 cells were obtained per trachea and showed up to 95% viability, as judged by trypan blue exclusion. When grown in culture, the cells formed monolayers after ~4 days. Electron microscopy of the monolayers revealed a polarized structure. An apical membrane, containing microvilli and a pronounced glycocalyx, was separated from a relatively unspecialized basolateral membrane by typical tight junctions. Monolayers grown on nucleopore filters showed resistances of 44-1,800 Ω·cm2 and transepithelial potential differences of 0.1-7.6 mV. Short-circuit current (I(sc)) was increased by isoproterenol, prostaglandins E2 and F(2α), and bradykinin. The loop diuretic, bumetanide, reduced I(sc) when added to the basolateral (serosal) side but had no effect from the apical (mucosal) side of the monolayers. Furosemide and MK-196 also inhibited I(sc). Mucosal amiloride inhibited I(sc). Serosal amiloride or mucosal ouabain had no effect on I(sc). Serosal ouabain brought I(sc) to zero after ~15 min.

Original languageEnglish (US)
Pages (from-to)1729-1735
Number of pages7
JournalJournal of Applied Physiology
Volume58
Issue number5
StatePublished - 1985
Externally publishedYes

ASJC Scopus subject areas

  • Endocrinology
  • Physiology
  • Orthopedics and Sports Medicine
  • Physical Therapy, Sports Therapy and Rehabilitation

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