CULTURED GASTRIC PARIETAL-CELLS FROM THE GUINEA-PIG - ADHERENCE, CELL-GROWTH AND STIMULUS COUPLING OF CA2-AMP( AND CYCLIC)

The aim of the study was to establish cell culture conditions for resp onsive guinea pig parietal cells. Parietal cells were isolated by a pr onase/collagenase method, enriched by counterflow elutriation and cult ured on plastic culture dishes in minimum essential medium. Precoating with gelatine or collagen increased adherence; optimum fetal calf ser um concentration was 10%. Parietal cells were cultured for up to 120 h . Intracellular calcium levels in cells cultured for 48 h were 150 nmo l/l and increased to 320 nmol/l after stimulation with carbachol and t o 250 nmol/l after histamine stimulation as determined by video imagin g microscopy. Intracellular cyclic AMP levels were increased 9-fold by histamine in cells cultured for 24 h and more than 30-fold in cells c ultured for 48 h. The results show that guinea pig parietal cells grow in primary culture and are suitable for studying second messenger cou pling.