ISOLATION, IDENTIFICATION AND QUANTITATIVE-EVALUATION OF SPECIFIC CELL-TYPES FROM THE MAMMALIAN GASTRIC-MUCOSA

Functional in vitro studies with isolated gastric mucosal cells requir e cytological identification of different cell types in suspension or primary culture. Since suitable techniques have not been well establis hed, different staining methods for the discrimination of dispersed pi g and guinea pig gastric cells have been developed on the basis of mod ified previous protocols for enzymatic cell dispersion. Chief and pari etal cells were visualized by combined periodic acid-Schiff stains. Su rface mucous and mucous neck cells were identified by affinity-labelli ng, using lectins with selective staining properties in situ. Two of t he lectins were found to be specific markers for gastric polymorphonuc lear cells. The following vital tests were found to be useful: succini c dehydrogenase for parietal cells, Nile blue/brilliant cresyl blue st ains for chief cells, and different phagocytosis assays for endothelia l cells and gastric phagocytes. Endocrine cells were characterized by immunocytochemistry using specific antibodies against gastrin, somatos tatin, histamine and serotonin. The same technique using a vimentin an tibody was performed for the identification of fibroblasts. Proliferat ion of mucosal cells in primary culture was monitored by the incorpora tion of bromo-deoxyuridine, which was subsequently detected by a monoc lonal antibody.