Cultured gastric parietal cells from the guinea pig: adherence, cell growth and stimulus coupling of Ca2+ and cyclic AMP.
The aim of the study was to establish cell culture conditions for responsive guinea pig parietal cells. Parietal cells were isolated by a pronase/collagenase method, enriched by counterflow elutriation and cultured on plastic culture dishes in minimum essential medium. Precoating with gelatine or collagen increased adherence; optimum fetal calf serum concentration was 10%. Parietal cells were cultured for up to 120 h. Intracellular calcium levels in cells cultured for 48 h were 150 nmol/l and increased to 320 nmol/l after stimulation with carbachol and to 250 nmol/l after histamine stimulation as determined by video imaging microscopy. Intracellular cyclic AMP levels were increased 9-fold by histamine in cells cultured for 24 h and more than 30-fold in cells cultured for 48 h. The results show that guinea pig parietal cells grow in primary culture and are suitable for studying second messenger coupling.
Giebel
J
J
Department of General Pharmacology, Medical School, Hannover.
Rechkemmer
G
G
Reimer
R
R
Sewing
K F
KF
Fanghänel
J
J
Schwenk
M
M
D000818:Animals; D002118:Calcium / Q000502:physiology; D002217:Carbachol / Q000494:pharmacology; D002448:Cell Adhesion; D002455:Cell Division; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000502:physiology; D005109:Extracellular Matrix / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D005753:Gastric Mucosa / Q000166:cytology*; D006168:Guinea Pigs; D017506:H(+)-K(+)-Exchanging ATPase / Q000378:metabolism; D010277:Parasympathomimetics / Q000494:pharmacology; D015398:Signal Transduction
D005353:Fibronectins; D010277:Parasympathomimetics; D002217:Carbachol; D000242:Cyclic AMP; D017506:H(+)-K(+)-Exchanging ATPase; D002118:Calcium