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Efficient and high-activity mouse intestinal epithelium and immune cell separation and extraction method

A technology for immune cells and intestinal epithelium, applied in cell dissociation methods, blood/immune system cells, gastrointestinal cells, etc., can solve problems such as cumbersome and time-consuming separation steps, low cell yield and activity, and limited cell research , to achieve the effect of simple method, high yield and high activity

Pending Publication Date: 2022-05-27
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods have cumbersome separation steps and take up to 6-10 hours, resulting in low cell yield and activity, which greatly limits subsequent research based on isolated cells

Method used

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  • Efficient and high-activity mouse intestinal epithelium and immune cell separation and extraction method
  • Efficient and high-activity mouse intestinal epithelium and immune cell separation and extraction method
  • Efficient and high-activity mouse intestinal epithelium and immune cell separation and extraction method

Examples

Experimental program
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Effect test

Embodiment

[0046] Example: A highly efficient and highly active method for separating and extracting mouse intestinal epithelium and immune cells, comprising the following steps:

[0047] Step 1, take the mouse small intestine (duodenum to the end of the ileum), wash it, cut it into pieces of 0.5-1cm, transfer the cut small intestine to a 50ml centrifuge tube, add the preheated temperature to 37 10ml of IEC separation solution at ℃, and then digested on a shaker for 15min at 37℃ and a rotating speed of 200rpm; the shaker is a metal bath shaker, and the maximum rotating speed can reach 400rpm;

[0048] Step 2, remove the 50ml centrifuge tube from the shaker, place it in a vortex vortex shaker for 15S, until the supernatant is chyle color, and separate intestinal epithelial cells and interepithelial immune cells;

[0049] Step 3, filter with a 100 μm membrane, take small intestine fragments, and wash the small intestine fragments with 2ml IEC separation solution; the filtrate after filter ...

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Abstract

The invention discloses an efficient and high-activity mouse intestinal epithelium and immune cell separation and extraction method, and belongs to the technical field of biological cell extraction. The intestinal epithelial cells, the interepithelial immune cells and the intestinal intrinsic layer immune cells separated and extracted by the method are high in yield and high in activity, and can be widely applied to related researches of intestinal mechanical barriers and immune barriers. Compared with the existing method, the separation and extraction steps are simple, the consumed time is greatly reduced, the single cell separation and purification can be completed within about 3.5 hours generally, and the subsequent research can be more efficiently carried out.

Description

technical field [0001] The invention belongs to the technical field of biological cell extraction, in particular to a method for separating and extracting mouse intestinal epithelium and immune cells with high efficiency and high activity. Background technique [0002] The gut is not only an important organ for the body to absorb nutrients, but also an important barrier for the body to fight microbial infections. In the process of intestinal anti-microbial infection, there are four protective barriers: microbial barrier, mucus barrier, mechanical barrier and immune barrier. The microbial barrier is mainly the normal flora of the human body, the mucous barrier is mainly composed of mucins and immunoglobulins secreted by the intestine, the mechanical barrier is mainly formed by the tight junctions between intestinal epithelial cells, and the immune barrier is composed of local intestinal immune cells and substances. . The gut is histologically divided into 4 layers: mucosa, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/078
CPCC12N5/0625C12N5/0679C12N5/0634C12N2509/10C12N2509/00
Inventor 梁廷波张匀张承陈诗音张剑谢雨婷俞文桥洪晓洁刘春晓傅淑蕾
Owner ZHEJIANG UNIV
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