Primary rat or mouse gastric mucosal epithelial cell isolation and culture method

An epithelial cell, separation and culture technology, applied in the field of cell culture, can solve the problems of large tissue cell damage, reduced cell yield and activity, and continuous in-depth research limitations, to improve separation, prevent blood clot adhesion, inhibit Effects of bacterial and fungal contamination

Inactive Publication Date: 2016-08-31
王晓冰
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  • Abstract
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Problems solved by technology

However, previous studies mostly used non-enzymatic mechanical separation methods or a single collagenase separation method.
The disadvantage is that the mechanical separation method and the single collagenase separation method will greatly damage the tissue cells, thereby greatly reducing the yield and activity of the cells, resulting in the separation and purification of gastric mucosal epithelial cells, which are not as easy to survive and grow as liver cells. , so that further continuous in-depth research is limited

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  • Primary rat or mouse gastric mucosal epithelial cell isolation and culture method
  • Primary rat or mouse gastric mucosal epithelial cell isolation and culture method

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Embodiment 1

[0039] In this embodiment, mice are used to separate gastric mucosal epithelial cells using the isolation and culture method of the present invention. The specific steps are as follows:

[0040] 1. Extraction of Mouse Stomach Tissue

[0041] Newborn or adult mice were first sacrificed (frozen for 10 minutes or cervical dislocated), soaked in alcohol for 5 minutes. Open the abdominal cavity to fully expose the abdominal cavity, and white gastric tissue can be seen. They were disconnected at the cardia and pylorus respectively, and the pylorus should be kept away from the pylorus when disconnecting, and gastric fundus tissue should be obtained as much as possible. Soak the removed gastric tissues in pre-cooled PBS containing 2% penicillin and streptomycin, and store them on ice. After all the tissues are collected, they will be processed uniformly.

[0042] 2. Washing and Preperfusion

[0043] After washing the isolated mouse or rat stomach tissue with 37°C preheated D-Hanks ...

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Abstract

The invention discloses a primary rat or mouse gastric mucosal epithelial cell isolation and culture method. A perfusion method is innovatively introduced into the primary rat or mouse gastric mucosal epithelial cell isolation and culture method, and sufficient perfusion without dead corners is achieved. Prepared compound enzyme digestive juice avoids the problems of low cell viability and poor yield and is capable of digesting more evenly and thoroughly as compared with preheated digestion of traditional single collagenase. Within 12 hours of cell inoculation adherence, a cell culture medium with a pH value of 6-6.5 is adopted, cell adherence rate can be increased greatly and adherence effects are better. The primary rat or mouse gastric mucosal epithelial cell isolation and culture method has the advantages of large total cell number, high isolation degree, high living rate and low contamination probability by bacteria, fungi and other cells as well as capability of achieving subculture so as to provide a favorable cell culture scheme for related fundamental researches.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a method for isolating and culturing primary rat or mouse gastric mucosal epithelial cells. Background technique [0002] Gastric mucosal epithelial cells have a high renewal rate, and various epithelial cells have different metabolic cycles, which are in a dynamic and delicate balance. The gastric mucosa is composed of many gastric units, and each gastric unit is composed of apical cells, parietal cells, chief cells, cervical mucous cells, pluripotent stem cells, and a small amount of endocrine cells and other epithelial cells. The apical cells, parietal cells, and chief cells are the three main types of cells in the gastric mucosa. Various epithelial cells are derived from pluripotent stem cells in the isthmus, each with different functions and turnover rates. Changes in cell differentiation, proliferation, and apoptosis directly affect the stability of gastric mucosal s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0679C12N2509/00
Inventor 王晓冰
Owner 王晓冰
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