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Method for utilizing gene chip to analyze lipopolysaccharide to stimulate variation of expression profile of dairy cow mammary epithelial cells

A mammary epithelial cell and gene expression profiling technology is applied in the field of analyzing the changes of the expression profile of dairy cow mammary epithelial cells stimulated by lipopolysaccharide by using a gene chip, and can solve the problems that standardization and mass production are difficult to overcome.

Inactive Publication Date: 2018-12-21
BEIJING UNIV OF AGRI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This method can greatly increase the lattice density, and the binding amount of each probe on the surface is relatively consistent, but there are still difficulties in standardization and mass production.

Method used

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  • Method for utilizing gene chip to analyze lipopolysaccharide to stimulate variation of expression profile of dairy cow mammary epithelial cells
  • Method for utilizing gene chip to analyze lipopolysaccharide to stimulate variation of expression profile of dairy cow mammary epithelial cells
  • Method for utilizing gene chip to analyze lipopolysaccharide to stimulate variation of expression profile of dairy cow mammary epithelial cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Embodiment 1, the establishment of model

[0078] Primary culture and isolation and purification of bovine mammary gland epithelial cells (BMECs)

[0079] Isolation and culture of BMECs

[0080] Select dairy cows in the lactation period without recessive mastitis, and obtain milk cow mammary gland tissue by aseptic surgical method; soak the mammary gland tissue in alcohol for 5 minutes, trim the tissue, and rinse repeatedly with distilled water until the liquid is clear; The acinar tissue was cut off and cleaned repeatedly, leaving no milk residue. Cut the mammary acini into 1mm3 tissue pieces, draw an appropriate amount of tissue pieces into a 15mL centrifuge tube, add 3 times the volume of 0.25% trypsin, shake and digest in a constant temperature water bath at 37°C for 30min, centrifuge at 1000r / min for 5min, and remove the supernatant , then add 3 times the volume of type II collagenase (0.2%), shake and digest in a constant temperature water bath for 1.5 hours at ...

Embodiment 2

[0105] Embodiment 2, detect Chinese medicine with model

[0106] Primary culture and isolation and purification of bovine mammary gland epithelial cells (BMECs)

[0107] Isolation and culture of BMECs

[0108] Select dairy cows in the lactation period without recessive mastitis, and obtain milk cow mammary gland tissue by aseptic surgical method; soak the mammary gland tissue in alcohol for 5 minutes, trim the tissue, and rinse repeatedly with distilled water until the liquid is clear; The acinar tissue was cut off and cleaned repeatedly, leaving no milk residue. Cut the mammary acini into 1mm3 tissue pieces, draw an appropriate amount of tissue pieces into a 15mL centrifuge tube, add 3 times the volume of 0.25% trypsin, shake and digest in a constant temperature water bath at 37°C for 30min, centrifuge at 1000r / min for 5min, and remove the supernatant , then add 3 times the volume of type II collagenase (0.2%), shake and digest in a constant temperature water bath for 1.5 h...

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Abstract

The invention relates to a method for utilizing a gene chip to analyze lipopolysaccharide to stimulate variation of the expression profile of dairy cow mammary epithelial cells. The method comprises the following steps that 1, a dairy cow mammary epithelial cell inflammation model is built up; and 2, traditional Chinese medicine is tested by using the dairy cow mammary epithelial cell inflammationmodel.

Description

Technical field: [0001] The present invention relates to drug screening and its curative effect detection method, in particular to a method for detecting lipopolysaccharide-stimulated milk cow mammary gland epithelial cell expression profile changes with a gene chip, so as to detect whether lipopolysaccharide can trigger an inflammatory response, and as an inflammation model to test the effect of drugs on inflammation The role of the model and the mechanism of action. Background technique: [0002] Traditional Chinese medicine refers to the natural medicine and its processing substitutes that are guided by the theory of traditional Chinese medicine, have a unique theoretical system and application form, are used to prevent and treat diseases, and have rehabilitation and health care functions, mainly including herbal medicine, animal medicine, and mineral medicine. . [0003] The therapeutic effect of traditional Chinese medicine has long been determined based on the change ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6837
CPCC12Q1/6837C12Q2521/107C12Q2565/501
Inventor 刘凤华李建东史雅然尹朋李焕荣侯晓林
Owner BEIJING UNIV OF AGRI
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