title: Gastric mucin secretion from cultured rat epithelial cells.
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.
authors.affiliation: Faculty of Pharmaceutical Sciences, and Life Science Research Center, Josai University, Sakado-shi, Saitama, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Tani
authors.forename: M
authors.initials: M
authors.lastname: Okuda
authors.forename: R
authors.initials: R
authors.lastname: Morishige
authors.forename: T
authors.initials: T
authors.lastname: Tanaka
mesh_terms: D000818:Animals; D002217:Carbachol / Q000494:pharmacology; D002478:Cells, Cultured; D015232:Dinoprostone / Q000494:pharmacology; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D005752:Gastric Mucins / Q000302:isolation & purification / Q000378:metabolism*; D005753:Gastric Mucosa / Q000187:drug effects / Q000378:metabolism*; D005755:Gastrins / Q000494:pharmacology; D006632:Histamine / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D012633:Secretin / Q000494:pharmacology; D012844:Sincalide / Q000494:pharmacology
chemical_list: D005752:Gastric Mucins; D005755:Gastrins; D012633:Secretin; D006632:Histamine; D002217:Carbachol; C036227:gastrin I; D015232:Dinoprostone; D012844:Sincalide