In vitro cell proliferation assay method using rat gastric cultured cells and effect of anti-ulcer drugs on the proliferation of cultured cells

Abstract

In order to establish a rapid and accurate in vitro drug assay method, we have applied the avidin D-biotin complex enzyme-linked immunosorbent assay using 5-bromodeoxyuridine (BrdU-ELISA) in the microplate cultures. To study the rat gastric cell proliferation, BrdU was treated with the cultured cells, and then, the rate of the uptake of BrdU into the cells was measured using an anti-BrdU antibody. Prostaglandin E2 (PGE2) (0.01, 0.1 and 1 microM), cimetidine (1, 3 and 10 microM) and omeprazole (3 and 10 microM) were shown to induce the significant proliferation of rat gastric cultured cells. Indomethacin (IND) (0.1, 0.25 and 0.5 mM) caused a dose-dependent inhibition of the proliferation of the cultured cells proliferation. The result obtained by the BrdU-ELISA method was more reproducible than that by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay method. PGE2 (0.01, 0.1 and 1 microM), cimetidine (1, 3 and 10 microM), omeprazole (1, 3 and 10 microM), cetraxate (3, 10 and 30 microM) and famotidine (30 microM), but not sofalcone and NC-1300-O-3, have been shown to protect significantly rat gastric cultured cells against IND-induced damage in vitro. From these results, it might be suggested that this BrdU-ELISA method is useful for investigating the effect of drugs on the proliferation of rat gastric cultured cells.