A rapid method for culturing guinea pig gastric mucous cell monolayers

D. W. Rattner, S. Ito, Michael Rutten, W. Silen

Research output: Contribution to journalArticle

33 Scopus citations

Abstract

A method has been developed for growing confluent primary cultured monolayers of guinea pig gastric mucous cells suitable for in vitro electrophysiological, transport, and pharmacological studies. Isolated mucous cells were enriched on a one-step Percoll density gradient and plated on fibronectin-coated plastic dishes or in small cups with holes containing glutaraldehyde-fixed Vitrogen gels. These cups were designed to fit in Ussing chambers. Mucous cells attached, proliferated, and formed confluent monolayers in 3 d. The low cuboidal cells contained periodic acid Schiff-positive mucous granules that were negative by Bowie and indirect immunofluorescent staining for pepsinogen. Electron microscopy revealed polarized mucous cells with microvilli, mucous granules, microfilaments, small mitochondria, some vacuoles, and junctional complexes that excluded wheat germ agglutinin-peroxidase. No basal lamina was present. Monolayers could be maintained for over 2 wk but subcultures were not made. The cultures were virtually free of fibroblasts. Epithelial sheets produced by this simple and rapid method can be used for electrophysiological, ion transport, and pharmacological studies.

Original languageEnglish (US)
Pages (from-to)453-462
Number of pages10
JournalIn Vitro Cellular & Developmental Biology
Volume21
Issue number8
DOIs
Publication statusPublished - Aug 1985
Externally publishedYes

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Keywords

  • cultured monolayer
  • gastric ion secretion
  • gastric mucous cell
  • primary cell culture

ASJC Scopus subject areas

  • Biotechnology
  • Plant Science
  • Cell Biology
  • Clinical Biochemistry
  • Developmental Biology

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