Primary culture of secretagogue-responsive parietal cells from rabbit gastric mucosa
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, (/sup 14/C)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.
- Research Organization:
- Morehouse School of Medicine, Atlanta, GA (USA)
- OSTI ID:
- 6499640
- Journal Information:
- Am. J. Physiol.; (United States), Vol. 256
- Country of Publication:
- United States
- Language:
- English
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ANIMAL CELLS
CYTOLOGICAL TECHNIQUES
ATP-ASE
BIOCHEMICAL REACTION KINETICS
CARBON 14 COMPOUNDS
CELL CULTURES
CELL MEMBRANES
CULTURE MEDIA
GASTROINTESTINAL TRACT
HISTAMINE
MONOCLONAL ANTIBODIES
PROTONS
RABBITS
TRACER TECHNIQUES
ULTRASTRUCTURAL CHANGES
ACID ANHYDRASES
AMINES
ANIMALS
ANTIBODIES
AZOLES
BARYONS
CELL CONSTITUENTS
DIGESTIVE SYSTEM
ELEMENTARY PARTICLES
ENZYMES
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HADRONS
HETEROCYCLIC COMPOUNDS
HYDROLASES
IMIDAZOLES
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
MAMMALS
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MORPHOLOGICAL CHANGES
NUCLEONS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOSPHOHYDROLASES
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550301* - Cytology- Tracer Techniques