A monolayer culture of gastric mucous cells from adult rabbits

Abstract

A new method for the primary monolayer cultures of adult rabbit gastric mucous cells has been developed. Rabbit gastric mucosal cells were isolated with etylenediaminetetraacetic acid and collagenase. Cells were cultured in Coon’s modified Ham’s F-12 medium supplemented with 10% fetal bovine serum, 15mM HEPES buffer, antibiotics, and antimycotic. The cells reached confluency on days 3–4. Histochemically 92% of the cells contained PAS positive gramules (mucous cells), 3% of cells showed a strong reaction for succinic dehydrogenase activity (parietal cells), 2% of the cells showed positive granules by Bowie staining (chief cells), and G6PDH staining was positive in 5% of the cells (surface mucous cells). Fibroblasts were rarely seen until day 7 (2 and PGI₂. Quantitatively cultured cells synthesized 1.475±0.039 ng/mg protein/hour of PGE₂ and 0.244±0.042 pg/mg protein/hour of PGI₂. This relatively simple and convenient technique provides a useful model for the study of cellular functions of gastric mucosa.