Method for culturing ruminal epithelial cells of sika deer

A technology of epithelial cells and sika deer, applied in cell dissociation methods, epidermal cells/skin cells, tissue culture, etc., can solve the problems of easy pollution, low vitality, and low purity of sika deer rumen epithelial cells, and achieve high purity and activity, overcoming the effects of pollution

Inactive Publication Date: 2018-06-01
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the problems of low purity, low vigor, low success rate and easy pollution of the above-mentioned cultured sika deer rumen epithelial cells, the present invention provides a method for culturing sika deer rumen epithelial cells

Method used

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  • Method for culturing ruminal epithelial cells of sika deer
  • Method for culturing ruminal epithelial cells of sika deer
  • Method for culturing ruminal epithelial cells of sika deer

Examples

Experimental program
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Effect test

Embodiment 1

[0032] Isolation and culture of sika deer rumen epithelial cells:

[0033] (1) Take the rumen tissue of sika deer, first wash the rumen content with normal saline, and then wash it with PBS buffer solution containing penicillin, streptomycin, gentamicin, and amphotericin B for 6 times, and wash it for the last time , then add PBS buffer and stand still for 10min;

[0034] (2) the muscular layer and the epithelial layer of the rumen tissue are separated, and the epithelial tissue is taken for subsequent use;

[0035] (3) Shred the epithelial tissue, spread the shredded tissue evenly in the culture bottle, place it upside down in the incubator for 2 hours;

[0036] (4) Add DMEM / F12 culture solution containing fetal bovine serum FBS, EGF, L-glutamine, insulin, transferrin, sodium selenite, penicillin, streptomycin, gentamicin, amphotericin B Carry out adherent culture;

[0037] After adding the culture medium, it was observed that cells crawled out after 3 days, and the number...

Embodiment 2

[0050] Isolation and culture of sika deer rumen epithelial cells:

[0051] (1) Take the rumen tissue of sika deer, first wash the rumen content with normal saline, and then wash it with PBS buffer solution containing penicillin, streptomycin, gentamicin, and amphotericin B for 8 times, and after the last wash , then add PBS buffer and stand still for 10min;

[0052] (2) the muscular layer and the epithelial layer of the rumen tissue are separated, and the epithelial tissue is taken for subsequent use;

[0053] (3) Shred the epithelial tissue, spread the shredded tissue evenly in the culture bottle, place it upside down in the incubator for 2 hours;

[0054] (4) Add DMEM / F12 culture solution containing fetal bovine serum FBS, EGF, L-glutamine, insulin, transferrin, sodium selenite, penicillin, streptomycin, gentamicin, amphotericin B Carry out adherent culture;

[0055] After adding the culture medium, it was observed that cells crawled out after 3 days, and the number of ce...

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Abstract

The invention provides a method for culturing ruminal epithelial cells of sika deer, relates to the field of isolation and culture of cells and aims to solve the problems of low purity, low vitality,low success rate, high probability of contamination and the like of culture of the ruminal epithelial cells of the sika deer. The method comprises the following steps: ruminal tissue of the sika deeris taken and washed with PBS containing antibiotics; epithelial tissue is taken and cut into pieces, and the pieces are added to a DMEM/F12 culture solution containing FBS (fetal bovine serum), sodiumselenite and the antibiotics for adherent culture; the pieces are added to the DMEM/F12 culture solution containing the FBS, sodium selenite and the antibiotics for subculture. The ruminal epithelialcells of the sika deer can be separated out successfully for the first time with the method, purity and activity are higher, steady passage can be realized, and the problem of contamination caused byvarious microorganisms in the rumen is solved. The method can be applied to research in growth and development, physiological functions, nutrition metabolism and other aspects of ruminal epithelium of the sika deer.

Description

technical field [0001] The invention relates to the field of isolating and culturing cells, in particular to cultivating sika deer rumen epithelial cells. Background technique [0002] Stomach and intestinal tract are the main digestive organs of animals, and rumen digestion is the main way for ruminants to digest feed. Therefore, the research on the nutrient transport and absorption mechanism of rumen epithelial cells has gradually become one of the hotspots of ruminant nutrition metabolism. The invention can be used for research on the growth and development, physiological function and nutrition metabolism of sika deer rumen epithelium. [0003] Primary culture is the only way to obtain rumen epithelial cells, and rumen epithelial cell lines are not sold in major cell banks at home and abroad. The rumen contains microorganisms such as bacteria, fungi, mycoplasma and protozoa. Therefore, the process of culturing cells is extremely susceptible to microbial contamination. A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0625C12N2509/00
Inventor 李志鹏南韦肖司华哲李光玉张宇飞
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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