Isolated culture method of rumen epithelial cells of adult yak

An epithelial cell, separation and culture technology, applied in cell dissociation methods, cell culture active agents, gastrointestinal cells, etc., can solve the problems reported in culturing adult yak rumen epithelial cells, not suitable for culturing adult yak rumen epithelial cells, etc. Mild effect, convenient use, cost-saving effect

Active Publication Date: 2021-03-09
SICHUAN AGRI UNIV
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Problems solved by technology

At present, the method of culturing rumen epithelial cells uses enzyme digestion, but the efficiency of collecting cells is related to the species, the size of the tissue block, the type of enzyme, the time of enzyme digestion, the way of enzyme digestion, the activity and concentration of the enzyme, and the living environment of yaks is special. , d

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  • Isolated culture method of rumen epithelial cells of adult yak
  • Isolated culture method of rumen epithelial cells of adult yak
  • Isolated culture method of rumen epithelial cells of adult yak

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Embodiment 1

[0030] Embodiment 1: a kind of isolation culture method of adult yak rumen epithelial cells, it comprises the following steps:

[0031] S1. Using sterilized scissors and tweezers, after the yak was sacrificed, the retroventral caeca tissue of the rumen was taken, and the rumen epithelial tissue with a size of 10 cm × 10 cm was cut, washed with tap water several times to remove the rumen content, and put into a beaker with 70% Rinse up and down with ethanol for 20 times, replace with new 70% ethanol and repeat this six times until it is cleaned, which can effectively remove residual bacteria and fungi; manually separate the epithelial and muscle layers with sterile gloves, and try to remove all Remove all the connective tissue, use sterilized scissors and tweezers to cut off all the edges of the epithelial layer, microorganisms will remain on the edge, and put the remaining tissue in a 50ml centrifuge tube and cut it into paste;

[0032] S2. First digest with 2 times the volume...

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Abstract

The invention discloses an isolated culture method of rumen epithelial cells of an adult yak. The method comprises the following steps of: collecting rumen epithelial tissues of the adult yak, cleaning the tissues with 70% absolute ethyl alcohol, and digesting the tissues step by combining I-type collagenase and trypsin; filtering and collecting cell filtrate through a nylon filter screen, and adding an MEM culture solution containing 10% fetal calf serum to terminate digestion; cleaning the collected cell precipitate; resuspending the cells by using a low-sugar DMEM complete medium, and theninoculating the cells into a culture flask for culture; and removing fibroblasts by a differential adherent method and a phase difference digestion method to obtain the rumen epithelial cells of the adult yak. According to the method provided by the invention, the rumen epithelial cells of the adult yak can be successfully separated, and the cells which are fast in adherent growth, relatively highin activity and stable in passage are obtained. The method can provide a test cell model for physiological regulation and nutrition absorption mechanism of the rumen epithelial cells of the yak.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for separating and culturing adult yak rumen epithelial cells. Background technique [0002] my country is a large yak breeding country in the world, and the stock of yaks accounts for more than 92% of the total yak breeding in the world. Yaks can adapt to the environment of high cold, low oxygen and lack of forage and become one of the dominant livestock species in the Qinghai-Tibet Plateau of China. They can provide meat and milk , fur and fuel (dung, as living fuel), play a vital role in the lives of local herders. And with the improvement of people's living standards, the consumption of yak meat continues to increase, and yak breeding has become a pillar industry of the pastoral economy. [0003] The rumen is an important organ for ruminant feed digestion and metabolism, nutrient absorption, nutrient deposition, and immune barrier. The volatile fatty acids p...

Claims

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Application Information

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IPC IPC(8): C12N5/071A01N1/02
CPCC12N5/0679A01N1/0226C12N2509/00C12N2501/11C12N2500/25C12N2500/30C12N2500/32Y02P60/87
Inventor 王之盛胡瑞王俊梅王雪莹姜雅慧代鹏
Owner SICHUAN AGRI UNIV
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