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Bovine rumen epithelial tissue dissociation method for single cell sequencing

A single-cell sequencing, epithelial tissue technology, applied in the field of cell biology, can solve the problem that the cell suspension cannot meet the requirements of single-cell sequencing

Active Publication Date: 2021-09-14
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Our previous research also found that the cell suspension obtained by the traditional rumen epithelial cell isolation and culture method in vitro cannot meet the requirements of single-cell sequencing, and further exploration is needed in the method

Method used

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  • Bovine rumen epithelial tissue dissociation method for single cell sequencing
  • Bovine rumen epithelial tissue dissociation method for single cell sequencing
  • Bovine rumen epithelial tissue dissociation method for single cell sequencing

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Embodiment Construction

[0037] The present invention will be described in detail below in conjunction with specific embodiments. The following examples will help those skilled in the art to further understand the present invention, but do not limit the present invention in any form. It should be noted that those skilled in the art can make several modifications and improvements without departing from the concept of the present invention. These all belong to the protection scope of the present invention.

[0038] A bovine rumen epithelial tissue dissociation method for single-cell sequencing provided by the present invention, such as Figure 8 As shown, the specific steps are:

[0039] (1) Sample mechanical treatment: put a sterile RNase-free cell culture dish on an ice box, add 1 × PBS; pick bovine rumen tissue and place it on a culture dish, wash it twice with 1 × DPBS, and observe the residue in the muscle layer case, use scissors and forceps to remove the muscle layer, and then cut the tissue b...

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Abstract

The invention discloses a bovine rumen epithelial tissue dissociation method for single cell sequencing. The bovine rumen epithelial tissue dissociation method comprises the following steps of: mechanically separating rumen tissues, performing enzyme digestion treatment, filtering impurities, cleaning cells, filtering again, observing the cells, controlling the quality, removing dead cells and the like. By adopting the bovine rumen epithelial tissue dissociation method, a rumen tissue single cell suspension with high activity, high cell count and low pollution can be obtained, the yield of effective single cell count and the quality of single cell nucleic acid are ensured, and a foundation is laid for single cell sequencing and data analysis; and the problem that a bovine rumen single cell dissociation technology is not systematically perfected in the prior art is solved.

Description

technical field [0001] The invention relates to the field of cell biology, in particular to a bovine rumen epithelial tissue dissociation method for single-cell sequencing. Background technique [0002] The rumen is a unique digestive organ of ruminants such as cows, which plays a vital role in the digestion, metabolism, absorption and transport of nutrients. Epithelial tissue is the main place where the rumen functions, but how to maintain and improve its structure and function in beef and dairy cattle farming still faces severe challenges, such as subacute rumen acidosis. Therefore, in-depth analysis of the key physiological functions and regulatory mechanisms of the rumen of dairy cows has become an important way to improve the health and production performance of dairy cows. Cells are the basic unit of life activities. The rumen epithelium is located on the innermost side of the rumen wall. It is a stratified squamous epithelium with four layers of multifunctional cell ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12Q1/6869
CPCC12N5/0679C12Q1/6869C12N2509/00C12N2509/10
Inventor 孙会增吴家劲刘潇菡朱森林顾凤飞刘建新
Owner ZHEJIANG UNIV
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