Primary culture of secretagogue-responsive parietal cells from rabbit gastric mucosa
| Autor: | C. S. Chew, A. Smolka, M. Ljungstrom, M. R. Brown |
|---|---|
| Rok vydání: | 1989 |
| Předmět: |
Male
Physiology Cell Separation Biology Basement Membrane H(+)-K(+)-Exchanging ATPase chemistry.chemical_compound Parietal Cells Gastric Physiology (medical) Gastrins medicine Animals Enterochromaffin-like cell Aminopyrine Cells Cultured Gastrin Parietal cell Adenosine Triphosphatases Hepatology Gastroenterology Culture Media Cell biology Kinetics Microscopy Electron Foveolar cell medicine.anatomical_structure chemistry Biochemistry Cell culture Carbachol Secretagogue Rabbits Protons G cell Cimetidine Histamine |
| Zdroj: | American Journal of Physiology-Gastrointestinal and Liver Physiology. 256:G254-G263 |
| ISSN: | 1522-1547 0193-1857 |
| DOI: | 10.1152/ajpgi.1989.256.1.g254 |
| Popis: | A new procedure for isolation and primary culture of gastric parietal cells is described. Parietal cells from rabbit gastric mucosa are enriched to greater than 95% purity by combining a Nycodenz gradient separation with centrifugal elutriation. Cells are plated on the basement membrane matrix, Matrigel, and maintained in culture for at least 1 wk. Parietal cells cultured in this manner remain differentiated, cross-react with monoclonal H+-K+-ATPase antibodies, and respond to histamine, gastrin, and cholinergic stimulation with increased acid production as measured by accumulation of the weak base, [14C]aminopyrine. When stimulated, cultured cells undergo ultrastructural changes in which intracellular canaliculi expand and numerous microvilli are observed. These ultrastructural changes are similar to those previously found to occur in vivo and in acutely isolated parietal cells. Morphological transformations in living cells can also be observed with differential interference contrast optics in the light microscope. After histamine stimulation, intracellular canaliculi gradually expand to form large vacuolar spaces. When the H2 receptor antagonist, cimetidine, is added to histamine-stimulated cells, these vacuoles gradually disappear. The ability to maintain hormonally responsive parietal cells in primary culture should make it possible to study direct, long-term effects of a variety of agonists and antagonists on parietal cell secretory-related activity. These cultured cells should also prove to be useful for the study of calcium transients, ion fluxes, and intracellular pH as related to acid secretion in single cells, particularly since morphological transformations can be used to monitor "physiological" responses at the same time within the same cell. |
| Databáze: | OpenAIRE |
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