Transepithelial transport by pulmonary alveolar type II cells in primary culture

R. J. Mason, M. C. Williams, Jonathan Widdicombe, M. J. Sanders, D. S. Misfeldt, L. C. Berry

Research output: Contribution to journalArticle

157 Scopus citations

Abstract

Fluid and electrolyte transport by epithelial cells in vitro can be recognized by the ability of cultured cells to form domes and by the electrical properties of monolayer cultures. Pulmonary alveolar epithelial cells are thought to be partially responsible for fluid movement in the fetal lung, but their role in electrolyte transport inthe adult lung is not known. We isolated alveolar type II cells from adult rat lung and maintained them on plastic culture dishes alone, on plastic culture dishes coated with an extracellular matrix, and on collagen-coated Millipore filters. Numerous large domes were formed on culture dishes coated with the extracellular matrix; smaller domes were formed on uncoated plastic culture dishes. Sodium butyrate (3 mM) stimulated dome formation. Transmission electron microscopy showed that the epithelial cells had flattened but still retained lamellar inclusions and that the cells were polarized with microvilli on the apical surface facing the culture medium. The electrical properties of the monolayers maintained on collagen-coated Millipore filters were tested in two laboratories. The transepithelial potential differences were 0.7 ± 0.1 mV (24 filters, seven experiments) and 1.3 ± 0.1 mV (13 filters, two experiments) apical side negative, and the corresponding resistances were 217 ± 11 ohm x cm 2 and 233 ± 12 ohm x cm 2. Terbutaline (10 μM) produced a biphasic response with a transient decrease and then a sustained increase in potential difference. Amiloride (0.1 mM) completely abolished the potential difference when it was added to the apical side but not when it was added to the basal side, whereas 1 mM ouabain inhibited the potential difference more effectively from the basal side. Thus, type II cells from a polarized epithelium in culture, and these cells actively transport electrolytes in vitro.

Original languageEnglish (US)
Pages (from-to)6033-6037
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume79
Issue number19 I
DOIs
StatePublished - 1982
Externally publishedYes

ASJC Scopus subject areas

  • Genetics
  • General

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