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Construction method for escherichia coli infected monolayer compact bovine mammary epithelial cell model

A technology of mammary gland epithelial cells and Escherichia coli, which is applied in the biological field and can solve problems such as endangering human and animal health and polluting the environment

Pending Publication Date: 2020-07-28
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the treatment of acute mastitis in cows mainly depends on the use of antibiotics, and the long-term and large-scale use of antibiotics has led to the emergence of drug resistance in many pathogenic bacteria, and the long-term and large-scale use of antibiotics will also pollute the environment and endanger human and animal health
At present, the damage mechanism of Escherichia coli-induced mastitis to mammary gland tissue and monolayer dense mammary epithelial cells has not been fully clarified, and the mechanism of many antibiotic substitutes to protect E. It is necessary to establish a monolayer dense dairy cow mammary epithelial cell model infected with Escherichia coli to provide a research basis

Method used

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  • Construction method for escherichia coli infected monolayer compact bovine mammary epithelial cell model
  • Construction method for escherichia coli infected monolayer compact bovine mammary epithelial cell model
  • Construction method for escherichia coli infected monolayer compact bovine mammary epithelial cell model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Construction and Application of Escherichia coli Infected Dairy Cow Monolayer Dense Mammary Epithelial Cell Model

[0064] 1. Materials

[0065] 1.1 Strains and cells

[0066] Escherichia coli strain number is ATCC 25922, provided by Beijing Zhongyuan Heju Biotechnology Co., Ltd.; cow mammary epithelial cells (Bovine mammary epithelial cells, BMECS), provided by Animal Nutrition and Feed Engineering Technology Research Center, College of Animal Science and Technology, Yangzhou University .

[0067] 1.2 Main reagents

[0068] The TSB and TSA medium used for strain culture were purchased from Beijing Suo Laibao Technology Co., Ltd.; the medium for cell culture, fetal bovine serum, etc. were purchased from Yingwei Jieji (Shanghai) Trading Co., Ltd.; the six-well plate for cell culture was purchased from Corning company; the rest of the analytical reagents were provided by the laboratory.

[0069] 1.3 Main instruments

[0070] CO 2 Constant temperature incub...

Embodiment 2

[0090] Example 2 Screening of Antibiotic Substitutes for Protecting Monolayer Dense Cow Mammary Epithelial Cell Damage

[0091] 1. Materials

[0092] 1.1 Strains and cells

[0093] Escherichia coli strain number is ATCC 25922, provided by Beijing Zhongyuan Heju Biotechnology Co., Ltd.; cow mammary epithelial cells (Bovine mammary epithelial cells, BMECS), provided by Animal Nutrition and Feed Engineering Technology Research Center, College of Animal Science and Technology, Yangzhou University ; Lactobacillus casei Zhang (Lactobacillus casei Zhang), provided by the Ministry of Education Key Laboratory of "Dairy Biotechnology and Engineering" of Inner Mongolia Agricultural University.

[0094] 1.2 Main reagents

[0095] The TSB and TSA medium used for strain culture were purchased from Beijing Suo Laibao Technology Co., Ltd.; the medium for cell culture, fetal bovine serum, etc. were purchased from Yingwei Jieji (Shanghai) Trading Co., Ltd.; the six-well plate for cell culture...

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Abstract

The invention provides a construction method for an escherichia coli infected monolayer compact bovine mammary epithelial cell model. The cell model is a monolayer compact bovine mammary epithelial cell model infected with escherichia coli is ATCC 25922, and during infection, the transepithelial electrical resistance value of the monolayer compact bovine mammary epithelial cell is 400 [omega]. cm<2>. The escherichia coli infected monolayer compact bovine mammary epithelial cell model provided by the invention can be used for detecting the adhesion rate of the escherichia coli infected monolayer compact bovine mammary epithelial cell and an influence of the escherichia coli infected monolayer compact bovine mammary epithelial cell for the expression of bovine mammary epithelial cell tight junction protein, and can be widely applied to related fields, including the screening of an injury mechanism of the escherichia coli induced mastitis for mammary tissues and mammary epithelial cells,and an antibiotic substitute.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for constructing a single-layer compact cow mammary gland epithelial cell model infected with Escherichia coli and its application. Background technique [0002] Mastitis is a kind of mammary gland inflammation caused by bacterial infection. It is one of the common and extremely harmful diseases in the dairy farming industry. It is a persistent disease in the global dairy farming industry. And other characteristics, not only caused great suffering to animals, but also brought huge economic losses to dairy farms all over the world. [0003] Escherichia coli is the main environmental pathogen that induces clinical mastitis in dairy cows, among which Escherichia coli ATCC25922 is an Escherichia coli strain isolated from mastitis cows, which can cause acute or hyperacute mastitis. [0004] At present, the treatment of acute mastitis in cows mainly depends on the use of antibiot...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N1/20C12Q1/02C12R1/19
CPCC12N5/0631C12N1/20G01N33/5008
Inventor 李胜利郑宇慧刘钢王蔚曹志军杨红建王雅晶
Owner CHINA AGRI UNIV
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