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Co-culture method capable of culturing anaerobic strains and pig intestine epithelial cells

A technology of co-cultivation of epithelial cells, applied to gastrointestinal cells, animal cells, vertebrate cells, etc., can solve the problems of dependence and inability to study bacteria in depth, achieve high success rate, simple operation process and equipment requirements, and improve experimental results. efficiency effect

Inactive Publication Date: 2021-06-11
SICHUAN AGRI UNIV
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Problems solved by technology

At present, amplicon sequencing or metagenomic sequencing is usually used to study the microbial composition and function of the gastrointestinal tract of animals, but these methods largely rely on bioinformatics methods, and the completeness of the reference database determines the accuracy of the analysis results. Correctness and precision, unable to study the specific role of a certain bacterium in depth from the aspects of physiology, metabolism, signal transduction, etc.

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  • Co-culture method capable of culturing anaerobic strains and pig intestine epithelial cells
  • Co-culture method capable of culturing anaerobic strains and pig intestine epithelial cells
  • Co-culture method capable of culturing anaerobic strains and pig intestine epithelial cells

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Embodiment 1

[0042] A method for cultivating anaerobic strains and porcine intestinal epithelial cells co-cultivates, comprising the following steps:

[0043] Step 1, adopt spectrophotometry to measure the optical density (OD) value of bacterial liquid:

[0044] Step 1.1. Preparation of anaerobic water: Measure 1L of distilled water and pour it into an Erlenmeyer flask, add 1mL of 0.1% resazurin solution at this time, the liquid turns blue; Disposable PE gloves and a rubber band seal the mouth of the Erlenmeyer flask; then pass the CO 2 or N 2 0.5~1h, add 1g of L-cysteine ​​or L-cysteine ​​hydrochloride, at this time the liquid is pink or yellow; finally ventilate again for 10min, seal and heat on low heat (do not boil) or seal and let stand for 10- After 12 hours, it can be observed that the medium becomes colorless or light yellow; the prepared anaerobic water is divided into well-airtight glass bottles for storage.

[0045] Among them, the preparation method of resazurin solution is...

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Abstract

The invention discloses a co-culture method for culturable anaerobic strains and pig intestine epithelial cells. The co-culture method comprises the following steps: measuring an optical density value of a bacterial solution by adopting a spectrophotometric method; determining the concentration of the bacterial liquid by adopting a plate counting method; determining the number of bacteria rapidly through correlation analysis of the two; and inoculating the accurately counted microorganisms to an intestinal epithelial cell line cultured in advance. According to the invention, the interference of various factors in other microorganisms or animal bodies can be eliminated, and the physiological functions of the microorganisms and the interaction mode of the microorganisms and host intestinal epithelial cells can be deeply analyzed.

Description

technical field [0001] The invention belongs to the field of micro-ecological nutrition and physiology, and in particular relates to a method for co-culturing cultivable anaerobic strains and pig intestinal epithelial cells. Background technique [0002] There are a large number of complex and diverse microbial flora inhabiting the intestinal tract of animals. A large number of studies have shown that intestinal microorganisms are closely related to the metabolism of nutrients, intestinal health and physiological conditions of the host. At present, amplicon sequencing or metagenomic sequencing is usually used to study the microbial composition and function of the gastrointestinal tract of animals, but these methods largely rely on bioinformatics methods, and the completeness of the reference database determines the accuracy of the analysis results. Correctness and precision, it is impossible to study the specific role of a certain bacterium in depth from the aspects of physi...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N5/071C12Q1/06C12R1/01
CPCC12N1/20C12N5/0679C12Q1/06G01N2333/195
Inventor 罗玉衡兰聪伍爱民李华谢坤宏田刚蔡景义陈代文
Owner SICHUAN AGRI UNIV
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