Post Date:Jun-02-23
As the uppermost part of the gastrointestinal tract, the stomach is an important digestive and metabolic organ. And the gastric epithelium and mucosa system is an essential maintainer of the physiological functions and pathological processes of the the stomach, which is a key research object for clinical and laboratory research of gastric diseases.
The stomach is an essential organ of the digestive system. It is a distensible organ between the esophagus and the duodenum, which can retain about 2-4L of food in adults[1-3]. One of the main functions of the stomach is food digestion. It can produce protease and hydrochloric acid to maintain an ideal environment (pH=2) for food digestion and harmful microorganism inhibition[4]. After digestion in the stomach, the food becomes chylous and then transported to the small intestine. The absorption of food is mainly located in the small intestine, but the stomach is responsible for the absorption of some small molecules, such as water, amino acids, alcohol (10-20%), some water-soluble vitamins, caffeine, and some drugs like aspirin[4-7]. Besides, the stomach is the controller of the whole progress of food digestion. The hormone secreted by the stomach and its peristalsis can control the speed of food entering the intestine. When the intestine is busy, the stomach will close itself and serve as a place for food storage[8].
The gastric epithelium and mucosa are part of the stomach wall that play irreplaceable roles in maintaining the function and structure of the stomach. And the diseases related to the stomach are mostly related to epithelium and mucosa[9, 10].(Figure.1) The stomach wall can withstand the acid environment in the cell conducive to digestion, while the gland is responsible for the secretion of enzymes, hormones, metabolic and immune factors, and acids[11]. The diseases or functional impairment of the stomach are mainly related to abnormal epithelial and mucosal, such as inflammation, cancer, etc., and harmful infections such as Helicobacter pylori, one of the leading causes of gastric epithelial and mucosal injury[11, 12].
Figure.1 Structure of stomach and gastric epithelium[11]
Gastric epithelial cells are base components of gastric epithelial, which maintain the physiological functions of the stomach. There are mainly four types of gastric epithelial cells: oxyntic (parietal) cells, zymogenic (chief) cells, surface mucous foveolar (pit) cells, and hormone-secreting enteroendocrine cells. These gastric epithelial cells form the wall and gland of the epithelial and continuous production of products involved in structural maintenance and food digestion and absorption. Besides, the self-renewal ability of these cells is also guaranteed for maintaining the structure and function of stomach organs[11]. Therefore, gastric epithelial cells are the key objectives of stomach physiology and pathology research.
In laboratory research of gastric physiology and diseases, in vitro models are valuable tools, mainly including primary gastric epithelial cells and immortalized gastric epithelial cell lines. Immortalized cell lines can have unlimited generations and are easy to culture and use. However, the immortalized transformation will inevitably change the physiological and pathological background, leading to the difference between the state of the cell and the actual organ. Therefore, the application of primary gastric epithelial cells is still common in high-impact publications. Primary gastric epithelial can provide a purer physiological background that makes in vitro models closer to actual in vivo processes, but the difficulties caused by its strict separation and cultivation make its application less convenient.
The isolation and culture protocol of human primary gastric cells will be briefly introduced[13]. The surgical gastric tissue must confirm without H. pylori and away from the tumor (the tissue resection position should be more than 10 cm from the tumor), and the tissue needs to be processed within 30 min after removal for the best results. After the sample is transferred to the operating environment in 4℃ RPMI 1640 medium containing antibiotics, a large amount of 4℃ PBS containing antibiotics is used to wash. And then, the mucosal layer is carefully removed from the muscle layer and cut into small pieces. The tissue fragments are digested and washed, continuing replenishing the digestive medium and taking out the supernatant containing multicellular lumps until the tissue is completely digested. And then the cell suspension was purified by PBS washing and centrifugation and then planked, and finally, the primary human cells were successfully obtained. After 24 hours of adherent culture, the cells are gently washed with PBS, and the culture medium is changed every 48 hours in the following culture.
Human gastric epithelial cells are widely used in research on the pathological mechanisms of gastric diseases caused by Helicobacter pylori. Morey et al. revealed a mechanism by which Helicobacter pylori escaped from the inflammatory reaction of gastric epithelial through primary human gastric epithelial cells and immortalized cell lines combined with mouse in vivo models[14]. They established in vitro Helicobacter pylori infection models on primary cells and MKN45, AGS cell lines, and treated the models with IFNγ and IFNβ. They found that cholesterol-α-glucosyltransferase(cgt) expressed by Helicobacter pylori can decrease the cholesterol level of the infected gastric epithelial cells to block the IFNγ signal through the JAK-STAT1 pathway, thereby allowing the bacteria to from the inflammatory reaction of the host.
Gastric diseases have been a hot research field for a long time, and the number of its publications has been at a high level in recent decades and is still rising. (Figure.2) It is essential for the researchers engaged in research in this field to master updated research technology at any time. Human gastric epithelial cells are valuable in vitro tools for the research of gastric physiology and pathology, and it is worth all researchers to be familiar with and master its application.
Figure.2 Publish data from NCBI PubMed
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[2] Curtis HNSB: Invitation to Biology. 1994.
[3] Wenzel V, Idris AH, Banner MJ, Kubilis PS, Band R, Williams JL, Jr., Lindner KH: Respiratory system compliance decreases after cardiopulmonary resuscitation and stomach inflation: impact of large and small tidal volumes on calculated peak airway pressure. Resuscitation 1998, 38:113-118.
[4] Levine. RMGMS: Textbook of Gastrointestinal Radiology. Philadelphia, PA.: Saunders.; 2007.
[5] Krehbiel CRM, J.C: Absorption of Amino acids and Peptides. Amino Acids in Animal Nutrition (2nd ed) 2015.
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[9] Stomach histology. Kenhub 2021.
[10] Dorland’s: Dorland’s Illustrated Medical Dictionary. Elsevier.; 2012.
[11] Mills JC, Shivdasani RA: Gastric epithelial stem cells. Gastroenterology 2011, 140:412-424.
[12] Li N, Xu X, Xiao B, Zhu E-D, Li B-s, Liu Z, Tang B, Zou Q-M, Liang H-P, Mao X-H: H. pylori related proinflammatory cytokines contribute to the induction of miR-146a in human gastric epithelial cells. Molecular Biology Reports 2012, 39:4655-4661.
[13] Qin J, Pei X: Isolation of Human Gastric Epithelial Cells from Gastric Surgical Tissue and Gastric Biopsies for Primary Culture. Methods Mol Biol 2018, 1817:115-121.
[14] Morey P, Pfannkuch L, Pang E, Boccellato F, Sigal M, Imai-Matsushima A, Dyer V, Koch M, Mollenkopf HJ, Schlaermann P, Meyer TF: Helicobacter pylori Depletes Cholesterol in Gastric Glands to Prevent Interferon Gamma Signaling and Escape the Inflammatory Response. Gastroenterology 2018, 154:1391-1404.e1399.
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