SUITABILITY OF DIFFERENT STAINING METHODS FOR THE IDENTIFICATION OF ISOLATED AND CULTURED-CELLS FROM GUINEA-PIG (CAVIA-APEREA-PORCELLUS) STOMACH

Cell suspensions from the guinea pig gastric mucosa were obtained usin g a pronase/collagenase isolation method, and cultured on Petri dishes in minimum essential medium at 37 degrees C. For proper identificatio n of different gastric cell types in cytospots, cell suspensions or cu lture, selective staining methods were employed, modified and evaluate d. Mucous cells and mucous neck cells were detected by use of lectins. Mucous cells were stained on cytospots and in primary cultures with l ectins from peanut, Helix pomatia, Illex europaeus, wheat germ, and fr om soybean. Vital chief cells in suspensions but not in culture, were selectively stained by Nile blue sulphate, brilliant cresyl blue or th e fluorescence dye dihexyloxacarbocyanine iodide. Pepsinogen granules of isolated and cultured chief cells were detected with a polyclonal a ntibody against porcine pepsinogen. Isolated parietal cells were ident ified in cytospots by using acidophilic dyes (aurantia, eosin). In sus pensions and in cultures vital parietal cells were identified by enzym atic detection of succinic dehydrogenase or carboanhydrase activity an d by the vital stain Janus green. In cultures exclusively, parietal ce lls were additionally identified by the vital stain rhodamine. Cytoche mically, they were identified with phalloidin by binding to actin fila ments. Endocrine cells in the suspension were visualised immunocytoche mically with antibodies directed against different amines or peptides. Fibroblasts and endothelial cells were identified after isolation and in primary culture with a vimentin antibody. Mast cells in suspension were either visualised by a histamine antibody or by metachromatic st aining behaviour to toluidine blue, respectively. Endothelial cells in suspension or culture were distinguished from fibroblasts by endocyto sis of acetylated low-density-lipoprotein. In conclusion, the develope d methods are highly suitable to identify guinea pig gastric cells aft er isolation and follow up their fate in primary culture.