Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:1.3.5.1 (succinate dehydrogenase)
 8,177 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The purpose of this study was to develop a primary cell culture system of rat gastric fundic epithelial cells. The cells, isolated enzymatically, were cultured in Coon's modified Ham's F-12 medium supplemented with 10% fetal bovine serum, 15 mM HEPES buffer, fibronectin, and antibiotics. The inoculated cells started to grow rapidly on day 1 (doubling time, 26 h). The cells reached confluency on day 3. On phase contrast microscopy, over 90% of cells possessed epithelial characteristics. Histochemical studies showed (a) 90% of the epithelial cells contained PAS positive granules, (b) 5% of the cells gave a strong reaction for succinic dehydrogenase activity (presumably parietal cells), and (c) immunohistochemical localization of pepsinogen was negative. Ultrastructurally, microvilluslike structures, junctional complexes, Golgi apparatus, mitochondria, rough-surfaced endoplasmic reticulum, and mucous granules were observed. Mitotic figures were clearly observed on Giemsa staining and the mitotic index was maximum on day 2. Autoradiographic and biochemical studies showed these cells possessed the capability to synthesize deoxyribonucleic acid and this ability was maximum on day 2. These cells were able to synthesize and to secrete glycoprotein and this function was significantly increased by 16,16-dimethyl prostaglandin E2. Cyclic adenosine monophosphate produced by the cultured cells was enhanced by addition of 16,16-dimethyl prostaglandin E2 (p less than 0.01). This in vitro system provides a valuable model for studies of cellular functions of gastric mucosa.
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PMID:Cell culture of rat gastric fundic mucosa. 629 Mar 9

Our aim was to develop a fibroblast-free monolayer culture of human gastric mucosal cells, using the specimens obtained by routine endoscopic biopsy. Human gastric mucosa obtained from normal volunteers by endoscopic biopsy was dissociated from collagenase and hyaluronidase. Dissociated cells were cultured in supplemented Coon's modified Ham's F-12 medium. Within 24 hr of inoculation, the cells were attached to the culture dishes. This was followed by cellular outgrowth. On phase-contrast microscopy, all cells had epithelial characteristics and fibroblasts were not observed. Ninety percent of cells contained periodic acid Schiff reaction-positive mucous granules after diastase digestion consistent with mucous epithelial cells. Two percent of the cells gave a strong reaction for succinic dehydrogenase activity (parietal cells). Immunohistochemical staining for pepsinogen in cultured cells was negative. On EM, microvilli-like projections, junctional complexes, Golgi apparatus, and mucous granules were apparent in the majority of cells. Mitotic figures were observed by day 3 with Giemsa staining. Autoradiographically, these cells were able to incorporate [3H]TdR into the nuclei. Cells were capable of synthesizing DNA, and this function was inhibited by cycloheximide. Cells could be cultured for up to two weeks without fibroblast contamination. A method of primary monolayer culture of human gastric mucosa obtained by a routine endoscopic biopsy has been successfully developed.
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PMID:A monolayer culture of human gastric epithelial cells. 686 89

Parietal cells of gastric glands are specialized to produce acid. Tight junctions between the parietal cells and their neighbouring cells (usually chief cells and mucous cells, less commonly parietal cells) avoid acid back-diffusion. Alterations of these junctions are accompanied by a defective epithelial barrier function. The conditions leading to junction formation, e.g. during epithelial restitution and regeneration are entirely unknown. The present study has the purpose to establish an in vitro model which allows studying these junctions. Freshly isolated gastric epithelial cells of guinea pig, moderately enriched with parietal cells, were cocultured for 2 days. Highly specific staining techniques showed the following composition in the near-confluent monolayer: 45% parietal cells (succinic dehydrogenase-positive), 36% mucous cells (lectin-binding granules), 18% chief cells (pepsinogen-positive granules) and 1% subepithelial cells (vimentin-positive). Ultrastructural investigations of sections of these monolayers revealed a high tendency of parietal cells to form cell junctions with the following characteristics: 1) virtually all parietal cells formed junctions with their neighbouring cells; 2) only junctions, but no desmosomes, were observed among neighbouring parietal cells; 3) junctional complexes and desmosomes were regularly present between parietal cells and their neighbouring mucous and chief cells; 4) parietal cells were sometimes integrated into three-dimensional structures, resembling rudimentary gastric glands. In conclusion, parietal cells under standard coculture conditions, generate de novo the same types of cell junctions that are observed in the intact gastric epithelium. The results show that parietal cells in vitro spontaneously make junctions with parietal and non-parietal cells, resembling the junctions in the intact tissue.
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PMID:Diversity of cell-cell interactions formed by gastric parietal cells in culture: morphological study on guinea pig cells. 840 32

Cell suspensions from the guinea pig gastric mucosa were obtained using a pronase/collagenase isolation method, and cultured on Petri dishes in minimum essential medium at 37 degrees C. For proper identification of different gastric cell types in cytospots, cell suspensions or culture, selective staining methods were employed, modified and evaluated. Mucous cells and mucous neck cells were detected by use of lectins. Mucous cells were stained on cytospots and in primary cultures with lectins from peanut, Helix pomatia, Ulex europaeus, wheat germ, and from soybean. Vital chief cells in suspensions but not in culture, were selectively stained by Nile blue sulphate, brilliant cresyl blue or the fluorescence dye dihexyloxacarbocyanine iodide. Pepsinogen granules of isolated and cultured chief cells were detected with a polyclonal antibody against porcine pepsinogen. Isolated parietal cells were identified in cytospots by using acidophilic dyes (aurantia, eosin). In suspensions and in cultures vital parietal cells were identified by enzymatic detection of succinic dehydrogenase or carboanhydrase activity and by the vital stain Janus green. In cultures exclusively, parietal cells were additionally identified by the vital stain rhodamine. Cytochemically, they were identified with phalloidin by binding to actin filaments. Endocrine cells in the suspension were visualised immunocytochemically 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 staining behaviour to toluidine blue, respectively. Endothelial cells in suspension or culture were distinguished from fibroblasts by endocytosis of acetylated low-density-lipoprotein. In conclusion, the developed methods are highly suitable to identify guinea pig gastric cells after isolation and follow up their fate in primary culture.
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PMID:Suitability of different staining methods for the identification of isolated and cultured cells from guinea pig (Cavia aperea porcellus) stomach. 883 36

We tested the hypothesis that recognized gastroprotective agents exert direct protection against ethanol-induced injury in isolated rat gastric mucosal cells in vitro. If protection exists, we also wanted to identify subcellular targets in the reversible and/or irreversible stages of cell injury. Ethanol-induced cell injury was quantified by measuring plasma membrane leakage (trypan blue exclusion and lactate dehydrogenase release), mitochondrial integrity (succinic dehydrogenase), and nuclear damage (ethidium bromide-DNA fluorescence). Initial cell viability and responsiveness were estimated by the effects of carbachol, carbachol + atropine, or 16,16-dimethyl-PGE(2) on chief cell pepsinogen secretion. Enriched parietal cells were stimulated by histamine, carbachol, or histamine + IBMX. Preincubation of cells with PG, sucrose octasulfate, or the sulfhydryl compounds N-acetylcysteine, taurine, or cysteamine increased cell resistance </=21% against ethanol. Similar protection was found with low histamine concentrations, but a higher concentration aggravated ethanol toxicity. Other naturally occurring or synthetic gastroprotective agents offered partial protection or aggravated ethanol-induced cell injury. Only a few in vivo gastroprotective agents demonstrated in vitro direct cytoprotection, which involved mainly the reversible stage of cell injury (e.g., plasma membrane changes) and, less often, irreversible (e.g., mitochondrial and nuclear) damage. Our findings also indicate that a major part of the beneficial effect of gastroprotective agents is expressed at the tissue level.
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PMID:Investigation of gastroprotective compounds at subcellular level in isolated gastric mucosal cells. 1109 42