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A method of co-culturing helicobacter pylori and gastric mucosa epithelial cells

A technology for Helicobacter pylori and epithelial cells, which is applied to gastrointestinal cells, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of physiological state influence, cumbersome bacterial work, etc., and achieve the effect of avoiding cumbersome

Inactive Publication Date: 2017-07-14
LANZHOU UNIVERSITY
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Problems solved by technology

[0003] The object of the present invention is exactly for the above-mentioned defect in the prior art, provides a kind of method for the co-cultivation of brand-new Helicobacter pylori and gastric mucosa epithelial cell, adds the sloping solid culture medium suitable for the growth of Helicobacter pylori in the normal cell culture bottle , so as to ensure the survival of Helicobacter pylori, and at the same time, the cell culture medium can ensure the normal growth of gastric mucosal cells. The impact of state differences and fluctuations in the number of viable bacteria on the experimental results, as well as the cumbersome work of adding bacteria multiple times

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  • A method of co-culturing helicobacter pylori and gastric mucosa epithelial cells
  • A method of co-culturing helicobacter pylori and gastric mucosa epithelial cells
  • A method of co-culturing helicobacter pylori and gastric mucosa epithelial cells

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Embodiment 1

[0024] A method for co-cultivating Helicobacter pylori and gastric mucosal epithelial cells, comprising the following steps:

[0025] 1) Prepare Columbia agar solid medium for cultivating Helicobacter pylori and sterilize it with high temperature and high pressure damp heat. When the solid medium is cooled to 50°C-60°C, add glucose solution and newborn calf serum, and mix well to obtain Helicobacter pylori medium; the preparation method of the Columbia agar solid medium for cultivating Helicobacter pylori is specifically: take 4.25g of Columbia agar base powder in a conical flask, add ddH 2 O 85ml, put into the bottle stopper and sterilize under high temperature and high pressure. When the solid medium is cooled to 50°C-60°C, add 5ml of 10% glucose injection so that the final concentration of glucose is 0.5% by mass volume ratio, and then Add 10ml of newborn calf serum so that the final concentration of newborn calf serum is 10% by volume, and mix well to obtain Helicobacter p...

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Abstract

A method of co-culturing helicobacter pylori and gastric mucosa epithelial cells is disclosed. The method includes 1) preparing a helicobacter pylori medium; 2) adding the medium into a sterile cell culture flask and preparing a slant surface; 3) culturing gastric mucosa epithelial cells in the culture flask in which the slant surface is formed and allowing the cells to be adhered to the wall of the flask; 4) centrifuging a helicobacter pylori 48-h culture liquid, and then resuspending a centrifuging product with a cell culture liquid; and 5) adding a resuspended bacterial liquid into the culture flask in which the slant surface and the adherent cells are formed, culturing, and counting the living bacteria and observing cell morphological changes every day. A beneficial effect of the method is that the novel method of co-culturing the helicobacter pylori and the gastric mucosa epithelial cells is provided. In the novel system, the helicobacter pylori can grow normally and continuously stimulate the adherent cells to generate lesions, thus simulating a human gastric mucosa infection situation well. The problem that helicobacter pylori declines or dies and cells are not effectively stimulated because culture liquids are not suitable for growth of the helicobacter pylori in traditional methods in which helicobacter pylori are directly added is overcome.

Description

technical field [0001] The invention relates to the technical field of bacteria and cell culture methods, in particular to a method for co-culture of Helicobacter pylori and gastric mucosal epithelial cells. Background technique [0002] Helicobacter pylori is a helical, Gram-negative microaerophilic bacterium discovered in 1983. It can colonize human gastric mucosal epithelial cells, causing gastritis, peptic ulcer, gastric cancer, gastric mucosa-associated lymphoid tissue lymphoma and other diseases. Since it is difficult to establish an animal model of Helicobacter pylori infection, the study of the pathogenic effect of Helicobacter pylori on cultured gastric mucosal epithelial cells in vitro is an important model for exploring the pathogenic mechanism of Helicobacter pylori. It is very important to establish a scientific and effective co-culture model of Helicobacter pylori and gastric mucosal epithelial cells. Since the pathogenicity of Helicobacter pylori on cells in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N1/20C12R1/01
CPCC12N1/20C12N5/0679C12N2502/70
Inventor 韩俭王礼彭琦孙旭冬任弟娟张富花徐媛媛
Owner LANZHOU UNIVERSITY
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