Isolated culture method for chicken enterocyte

Abstract

The invention relates to an isolated culture method for chicken enterocyte. The method comprises the following steps: step 1, digesting the intestinal tissue block of a chicken embryo with protease so as to obtain a digestion product; step 2, filtering the digestion product with a cell strainer and collecting cell aggregate with a size of 100 to 500 meshes; and step 3, subjecting the cell aggregate to centrifugation at a speed of 800 to 1500 r / min for 5 to 15 min and collecting cell sediment. The invention further provides the chicken enterocyte obtained by using the method. The method provided by the invention can substantially shorten testing time, save reagents and improve the purity of the isolated enterocyte.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for separating and culturing chicken intestinal epithelial cells. Background technique [0002] Intestinal epithelial cells are the medium of the environment inside and outside the intestinal tract, and are also an important part of the body's immune barrier, with important physiological functions such as digestion, absorption, secretion, and immunity. Intestinal epithelial cells can be isolated and cultured in vitro under certain conditions. Isolation and culture of intestinal epithelial cells in vitro provides a simple, rapid and accurate means for the study of intestinal biological functions, nutrient absorption mechanism and its regulation, as well as the proliferation, differentiation and apoptosis of intestinal epithelial cells. The role of the intestinal epithelium provides an ideal in vitro model. However, the isolation and culture of intestinal epithelial cells is...

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Problems solved by technology

However, there are certain differences between intestinal cancer epithelial cells and normal epithelial cells in terms of cell basic structure, metabolism, and functional characteristics, and there are great limitations in explaining normal behaviors with the research results of cancer cell lines

[0003] At present, there is no patent document about the in vitro isolation or culture method of primary chicken intestinal epithelial cells. Although conventional cell culture techniques are used in the existing chicken intestinal epithelial cell culture methods, the purpose of culture can be achieved to a certain extent, but there are The operation is cumbersome, the cells are not easy to adhere to the wall, the growth is slow, and other miscellaneous cells such as fibroblasts are difficult to separate, etc., which cannot meet the requirements of cell experiments

[0004] The inventors of the present invention have found through long-term experimental research that the growth of intestinal epithelial cells mixed with fibroblasts is often due to the inability to separate intestinal epithelial cells and fibroblasts when separating cells. Isolation of fibroblast components, however the process is tedious and time consuming, typically 5-7 hours and the cell purity is low

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