Isolation, culture and adenoviral transduction of parietal cells from mouse gastric mucosa

Biomedical Materials
Briony L GliddonIan R van Driel

Abstract

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 an acid-secreting active parietal cell were observed. In resting cultures of mouse parietal cells, the H(+)/K(+) ATPase displayed a cytoplasmic punctate staining pattern consistent with tubulovesicle element structures. Following histamine stimulation, an expansion of internal apical vacuole structures was observed together with a pronounced redistribution of the H(+)/K(+) ATPase from the cytoplasm to the apical vacuoles. A reproducible procedure to express genes of interest exogenously in these cultures of mouse parietal cells was also estab...Continue Reading

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Citations

Apr 1, 2011·Therapeutic Delivery·Nicolas LaurenttOlivier Zelphati
Dec 12, 2013·Molecular Metabolism·Maja S EngelstoftThue W Schwartz
Oct 21, 2016·Veterinary Research·Guangzhi ZhangFreddy Haesebrouck
Nov 17, 2012·American Journal of Physiology. Gastrointestinal and Liver Physiology·Nhung NguyenIan R van Driel
Jun 21, 2017·Biomedical Materials·Yakup UlusuDaniel T Peters
Mar 30, 2017·Proceedings of the National Academy of Sciences of the United States of America·Katsuhiro KawaaiKatsuhiko Mikoshiba

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