Gastric mucin secretion from cultured rat epithelial cells

Biological & Pharmaceutical Bulletin
S TaniT Tanaka

Abstract

In order to establish the measurement of gastric mucin secreted from cultured mucous cells, rat gastric mucin was purified from secreted mucus with Sepharose CL-4B column chromatography. Gastric mucin was measured by dot blot analysis using an enzyme-linked lectin (soybean agglutinin) assay in a good concentration-dependent manner. Surface epithelial cells were dispersed by limited digestion of a rat everted stomach and collected by density gradient centrifugation with Percoll. These cells were inoculated onto gelled collagen dishes, then cultured in a medium supplemented with 10% fetal calf serum under a 5% CO2 atmosphere in air. Changing the medium after a 2-d culture, the cells were cultured for another 3 d. During the culture, the numbers of cells each day were almost equal, but mucin contents in the cells increased, and then dropped at day 5 after inoculation. At that time, the edge of the cell layer peeled off and the cells adhered to each other. Using 2-d cultured cells, the effects of some secretagogues on mucin secretion were investigated. Carbachol, secretin, CCK-8 and prostaglandin E2 (PGE2) strongly stimulated mucin secretion, and gastrin I weakly did. However, histamine offered no stimulation.

Citations

Aug 29, 2006·PLoS Biology·K MiyakeP L McNeil
Oct 4, 2006·British Journal of Pharmacology·I DeyK Chadee
Aug 26, 2006·Regulatory Peptides·Francis K Y SiuBilly K C Chow
Apr 30, 2011·Pharmacological Research : the Official Journal of the Italian Pharmacological Society·Pratibha SinghGautam Palit
Oct 3, 2017·Biological & Pharmaceutical Bulletin·Mayumi TamuraYoichiro Arata
May 11, 2004·Biological & Pharmaceutical Bulletin·Satoru TaniToru Tanaka
Feb 6, 2020·Biological & Pharmaceutical Bulletin·Mayumi TamuraYoichiro Arata

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