Isolation, culture and adenoviral transduction of parietal cells from mouse gastric mucosa
Briony L Gliddon, Nhung V Nguyen, Priscilla A Gunn, Paul A Gleeson, Ian R van Driel
BIOMEDICAL MATERIALS | IOP PUBLISHING LTD | Published : 2008
Here we describe a method for the isolation of intact gastric glands from mice and primary culture and transfection of mouse gastric epithelial cells. Collagenase digestion of PBS-perfused mouse stomachs released large intact gastric glands that were plated on a basement membrane matrix. The heterogeneous gland cell cultures typically contain approximately 60% parietal cells. Isolated mouse parietal cells remain viable in culture for up to 5 days and react strongly with an antibody specific to the gastric H(+)/K(+) ATPase. Isolated intact mouse gastric glands and primary cultures of mouse parietal cells respond to the secretagogue, histamine. Typical morphological changes from a resting to a..View full abstract
The authors thank Catherine Chew from the Institute of Molecular Medicine and Genetics at the Medical College of Georgia for helpful technical advice. Dr Sarah Londrigan from the Walter and Eliza Hall Institute is greatly acknowledged for helpful advice on the production of recombinant adenovirus. This work was supported by funding from the Australian Research Council.