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Method for constructing non-infectious gastric mucosal lesion cell model and application

A technology for gastric mucosal injury and cell models, which is applied in the direction of gastrointestinal cells, animal cells, vertebrate cells, etc., can solve the problems of cumbersome operation, difficult large-scale sample screening, large demand for experimental samples, etc., and achieve good model stability , short modeling time and easy operation

Pending Publication Date: 2022-03-01
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In recent years, polysaccharides such as Hericium erinaceus and Dendrobium have also been found to have protective effects on gastric mucosal injury or gastric ulcer in animal models. However, due to the long experimental cycle of animal models, the operation is cumbersome and involves animal ethics issues. At the same time, the experimental sample requirements Large amount, difficult to screen large batches of samples
Existing studies also lack the evaluation of the gastric mucosal protective activity of polysaccharide samples from different sources in the same model

Method used

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  • Method for constructing non-infectious gastric mucosal lesion cell model and application
  • Method for constructing non-infectious gastric mucosal lesion cell model and application
  • Method for constructing non-infectious gastric mucosal lesion cell model and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Take out the cell cryopreservation tube from the liquid nitrogen tank, quickly place it in a 37°C water bath to dissolve until there are no solid crystals, wipe the outer wall with 75% alcohol, and transfer it to a medium containing 5ml of complete medium (containing 10% FBS). DMEM medium) in a 15ml sterile centrifuge tube, blow and mix with a pipette gun, centrifuge at 1000rmp for 5min. Discard the supernatant, collect the precipitated cells, add 1ml of complete medium to break up the cells, and place at 37°C, 5% CO 2 Cell culture in the incubator box.

[0032] When the cell density in the culture flask reaches about 80%, suck off the old culture medium, gently wash it twice with PBS, add 2ml of 0.25% trypsin digestion solution, digest it in a 37°C incubator for 2min, take it out and examine it under the microscope Next, observe the digestion of the cells. If most of the cells become round and scattered, add 5ml of complete medium to stop the digestion. Centrifuge at...

Embodiment 2

[0035] The difference with Example 1 is that the ethanol concentration is 6%, and all the other are the same as Example 1.

Embodiment 3

[0037] The difference with Example 1 is that the ethanol concentration is 7%, and all the other are the same as Example 1.

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Abstract

The invention provides a method for constructing a non-infectious gastric mucosal lesion cell model and application, and belongs to the technical field of cell model construction. The method for constructing the non-infectious gastric mucosal lesion cell model comprises the following steps: taking 2 * 10 < 4 >-2 * 10 < 5 > cells / mL logarithmic phase GES-1 cells for conventional culture, adding a culture solution containing ethanol or H2O2 after the cells are adhered to the wall, culturing for 6-24 hours, and discarding the lesion solution, thereby obtaining the induced gastric mucosal lesion cell model. Two exogenous attack factors of ethanol and H2O2 are used for inducing human gastric mucosal epithelial cells to construct the gastric mucosal lesion model, compared with an animal model, the cell model is easier to obtain and simple and convenient to operate, high-throughput screening of samples can be achieved, the method is short in modeling time, the constructed model is good in stability, and the method is suitable for large-scale popularization and application. The method can be applied to research and development of stomach protecting products or research of non-infectious gastric mucosal lesion mechanisms.

Description

technical field [0001] The invention belongs to the technical field of cell model construction, and in particular relates to a method and application for constructing a non-infectious gastric mucosal injury cell model. Background technique [0002] The stomach is an organ that plays a digestive role in the human body. The dynamic balance between its mucosal layer attack factors and mucosal self-defense repair factors is the key to ensuring the normal work of the stomach. Once this dynamic balance is broken, the gastric acid produced will gradually damage the stomach wall. Cause mucosal damage, cause gastritis, gastric ulcer and other diseases related to gastric mucosal injury. With the progress of society and the times, the acceleration of the pace of life, and the influence of bad living habits, the incidence of stomach diseases has shown a rapid increase trend. Gastric ulcer has also become a common digestive disease in internal medicine and is currently the most serious d...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12Q1/02
CPCC12N5/0679G01N33/5044C12N2500/30C12N2500/05
Inventor 吴迪杨焱马强张忠张劲松陈万超李文刘艳芳
Owner SHANGHAI ACAD OF AGRI SCI
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