A collagen IV matrix is required for guinea pig gastric epithelial cell monolayers to provide an optimal model of the stomach surface for biopharmaceutical screening

Klairi M. Kavvada, James G. Murray, Vanessa A. Moore, Alan G.A. Coombes, Peter J. Hanson

Research output: Contribution to journalArticle

Abstract

The surface epithelial cells of the stomach represent a major component of the gastric barrier. A cell culture model of the gastric epithelial cell surface would prove useful for biopharmaceutical screening of new chemical entities and dosage forms. Primary cultures of guinea pig gastric mucous epithelial cells were grown on filter inserts (Transwells®) for 3 days. Tight-junction formation, assessed by transepithelial electrical resistance (TEER) and permeability of mannitol and fluorescein, was enhanced when collagen IV rather than collagen I was used to coat the polycarbonate filter. TEER for cells grown on collagen IV was close to that obtained with intact guinea pig gastric epithelium in vitro. Differentiation was assessed by incorporation of [ 3H]glucosamine into glycoprotein and by activity of NADPH oxidase, which produces superoxide. Both of these measures were greater for cells grown on filters coated with collagen I than for cells grown on plastic culture plates, but no major difference was found between cells grown on collagens I and IV. The proportion of cells, which stained positively for mucin with periodic acid Schiff reagent, was greater than 95% for all culture conditions. Monolayers grown on membranes coated with collagen IV exhibited apically polarized secretion of mucin and superoxide, and were resistant to acidification of the apical medium to pH 3.0 for 30 min. A screen of nonsteroidal anti-inflammatory drugs revealed a novel effect of diclofenac and niflumic acid in reversibly reducing permeability by the paracellular route. In conclusion, the mucous cell preparation grown on collagen IV represents a good model of the gastric surface epithelium suitable for screening procedures. © 2005 The Society for Biomolecular Screening.

Original languageEnglish
Pages (from-to)495-507
Number of pages13
JournalJournal of Biomolecular Screening
Volume10
Issue number5
DOIs
Publication statusPublished - Aug 2005

Fingerprint

Monolayers
Stomach
Screening
Guinea Pigs
Collagen
Epithelial Cells
Cells
polycarbonate
Acoustic impedance
Mucins
Electric Impedance
Cell culture
Superoxides
Permeability
Epithelium
Niflumic Acid
Periodic Acid
Acidification
Tight Junctions
Diclofenac

Keywords

  • gastric
  • mucous cell
  • nonsteroidal anti-inflammatory drug
  • stomach
  • Transwells®

Cite this

Kavvada, Klairi M. ; Murray, James G. ; Moore, Vanessa A. ; Coombes, Alan G.A. ; Hanson, Peter J. / A collagen IV matrix is required for guinea pig gastric epithelial cell monolayers to provide an optimal model of the stomach surface for biopharmaceutical screening. In: Journal of Biomolecular Screening. 2005 ; Vol. 10, No. 5. pp. 495-507.
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abstract = "The surface epithelial cells of the stomach represent a major component of the gastric barrier. A cell culture model of the gastric epithelial cell surface would prove useful for biopharmaceutical screening of new chemical entities and dosage forms. Primary cultures of guinea pig gastric mucous epithelial cells were grown on filter inserts (Transwells{\circledR}) for 3 days. Tight-junction formation, assessed by transepithelial electrical resistance (TEER) and permeability of mannitol and fluorescein, was enhanced when collagen IV rather than collagen I was used to coat the polycarbonate filter. TEER for cells grown on collagen IV was close to that obtained with intact guinea pig gastric epithelium in vitro. Differentiation was assessed by incorporation of [ 3H]glucosamine into glycoprotein and by activity of NADPH oxidase, which produces superoxide. Both of these measures were greater for cells grown on filters coated with collagen I than for cells grown on plastic culture plates, but no major difference was found between cells grown on collagens I and IV. The proportion of cells, which stained positively for mucin with periodic acid Schiff reagent, was greater than 95{\%} for all culture conditions. Monolayers grown on membranes coated with collagen IV exhibited apically polarized secretion of mucin and superoxide, and were resistant to acidification of the apical medium to pH 3.0 for 30 min. A screen of nonsteroidal anti-inflammatory drugs revealed a novel effect of diclofenac and niflumic acid in reversibly reducing permeability by the paracellular route. In conclusion, the mucous cell preparation grown on collagen IV represents a good model of the gastric surface epithelium suitable for screening procedures. {\circledC} 2005 The Society for Biomolecular Screening.",
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A collagen IV matrix is required for guinea pig gastric epithelial cell monolayers to provide an optimal model of the stomach surface for biopharmaceutical screening. / Kavvada, Klairi M.; Murray, James G.; Moore, Vanessa A.; Coombes, Alan G.A.; Hanson, Peter J.

In: Journal of Biomolecular Screening, Vol. 10, No. 5, 08.2005, p. 495-507.

Research output: Contribution to journalArticle

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AU - Kavvada, Klairi M.

AU - Murray, James G.

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AU - Coombes, Alan G.A.

AU - Hanson, Peter J.

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