IN-VITRO CELL-PROLIFERATION ASSAY-METHOD USING RAT GASTRIC CULTURED-CELLS AND EFFECT OF ANTIULCER DRUGS ON THE PROLIFERATION OF CULTURED-CELLS

In order to establish a rapid and accurate in vitro drug assay method, we have applied the avidin D-biotin complex enzyme-linked immunosorbe nt assay using 5-bromodeoxyuridine (BrdU-ELISA) in the microplate cult ures. To study the rat gastric cell proliferation, BrdU was treated wi th the cultured cells, and then, the rate of the uptake of BrdU into t he cells was measured using an anti-BrdU antibody. Prostaglandin E2, ( PGE2) (0.01, 0.1 and 1 mum), cimetidine (1, 3 and 10 mum) and omeprazo le (3 and 10 mum) were shown to induce the significant proliferation o f rat gastric cultured cells. Indomethacin (IND) (0.1, 0.25 and 0.5 mm ) caused a dose-dependent inhibition of the proliferation of the cultu red cells proliferation. The result obtained by the BrdU-ELISA method was more reproducible than that by the (4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay method. PGE2 (0.01, 0.1 and 1 mum), cimetidine (1, 3 and 10 mum), omeprazole (1, 3 and 10 muM), cetr axate (3, 10 and 30 mum) and famotidine (30 mum), but not sofalcone an d NC-1300-0-3, have been shown to protect significantly rat gastric cu ltured cells against IND-induced damage in vitro. From these results, it might be suggested that this BrdU-ELISA method is useful for invest igating the effect of drugs on the proliferation of rat gastric cultur ed cells.