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

Abstract

The surface epithelial cells of the stomach represent a major component of the gastric barrier. Acell culturemodel of the gastric epithelial cell surface would prove useful for biopharmaceutical screening of new chemical entities and dosage forms. Primary cultures of guinea pig gastricmucous epithelial cellswere grown on filter inserts (Transwells®) for 3 days. Tight-junction formation, assessed by transepithelial electrical resistance (TEER) and permeability ofmannitol 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 coatedwith collagen I than for cells grown on plastic culture plates, but nomajor 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, themucous cell preparation grown on collagen IV represents a good model of the gastric surface epithelium suitable for screening procedures.