K. Imai et al., IN-VITRO CELL-PROLIFERATION ASSAY-METHOD USING RAT GASTRIC CULTURED-CELLS AND EFFECT OF ANTIULCER DRUGS ON THE PROLIFERATION OF CULTURED-CELLS, Yakugaku zasshi, 114(5), 1994, pp. 316-324
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.