Isolated culture method of rumen epithelial cells of adult yak

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...

AI Technical Summary

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. , different from other cattle, and adult yak rumen epithelial cells are already in an aging state. Therefore, the method for obtaining rumen epithelial cells in existing patent documents is not suitable for culturing adult yak rumen epithelial cells. After searching, the prior art does not There is a report about culturing adult yak rumen epithelial cells

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