An organ culture method for glandular stomach of new born rat.

Abstract

An organ culture system was developed for the long-term maintenance of the glandular stomach of newborn Wistar rats. The explants from glandular stomach were cultured on Millipore filter in an organ culture dish with various media consisting of sera, buffers and additives under different gas phases of O2, CO2 and N at 37 or 24.degree. C. The culture condition was evaluated by histological examination of explants and was divided into Grade (+++), (++), (+) and (-) depending upon the morphology and growth of the mucosal epithelium. D-MEM [Eagles'' minimum essential medium] plus fetal calf serum (20%) supplemented with hydrocortisone (10 .mu.g/ml) or dexamethasone (5 .mu.g/ml) buffered with sodium bicarbonate and Hepes under a gas phase of O2 (95%) + CO2 (5%) or O2 (45%) + CO2 (5%) + N2 (50%) at 37.degree. C showed the best result. Other supplements tested, such as insulin, ascorbic acid and ferrous sulfate were ineffective. Addition of hydrocortisone or dexamethasone was similarly effective in serum-free media. Isologous rat serum had no advantage over fetal calf serum. Transplantation and autoradiography revealed the viability of the explants up to 35 days in vitro.