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Method for culturing airway epithelial cells by simply, conveniently and rapidly inducing differentiation of cilia

An airway epithelial cell and culture method technology, which is applied in the field of airway epithelial cell culture, can solve the problems of long airway epithelial cell culture period, many tools, and small amount of cells, and achieves accelerated growth and collection speed and simple technical process. Fast, high-quality cell culture results

Inactive Publication Date: 2016-03-30
李童斐 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of the deficiencies occurring in the above-mentioned method, the purpose of the present invention is to overcome the deficiencies in the above-mentioned technical method and propose a solution, to eliminate the airway epithelial cell culture cycle is long, the operation process is complicated, and there are many tools to be used. The amount of cells produced is small, especially to eliminate the problem of poor differentiation of cultured epithelial cells that leads to less or no cilium swinging of airway epithelial cells, and provides a simple and rapid method for culturing airway epithelial cells that can induce ciliated differentiation

Method used

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  • Method for culturing airway epithelial cells by simply, conveniently and rapidly inducing differentiation of cilia
  • Method for culturing airway epithelial cells by simply, conveniently and rapidly inducing differentiation of cilia
  • Method for culturing airway epithelial cells by simply, conveniently and rapidly inducing differentiation of cilia

Examples

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Effect test

Embodiment 1

[0022] A. In a sterile room temperature environment, the SPF grade SD rats grown to 2 weeks old were inhaled anesthetized, and the neck skin of the SD rats was cut with surgical scissors, and the fascia under the skin was torn off with surgical forceps to expose Out of the airway, a section of airway tissue is isolated.

[0023] B. Rinse the separated airway tissue in a petri dish containing phosphate buffered saline at room temperature until the airway does not contain mucus and other impurities.

[0024] C. Lay the rinsed airway tissue horizontally, and cut it longitudinally into six independent annular airway tissues, each with a thickness of about 1 mm.

[0025] D. Use pointed forceps to carefully clamp the independent ring tissues, and be careful not to damage the inner side of the airway ring. Spread the ring tissues in 6 wells of a 6-well plate. One hole in the plate was covered with slides, and after the airway ring was placed, 500 μl of medium was slowly added to eac...

Embodiment 2

[0028] A. Under a sterile room temperature environment, decapitate SPF C57 mice grown to the age of 2-5 weeks, use surgical scissors to open the chest cavity of C57 mice in a sterile workbench, and separate the trachea, main bronchi and lungs. Take out the tissue together.

[0029] B. Separate the removed trachea and main bronchi from the lung tissue, place the trachea and main bronchi in phosphate buffered saline after a water bath containing penicillin and streptomycin at 37 degrees Celsius.

[0030] C. Use surgical scissors to cut the rinsed airway tissue into 3 independent columnar trachea, place the 3 columnar trachea horizontally, and use a scalpel to cut the columnar trachea longitudinally into 15 tracheal rings with a thickness of 1 mm.

[0031] D. Use pointed forceps to place 15 circular airway tissues horizontally in the same culture well of a 6-well plate, with an interval of 2 to 5 mm between the 15 circular airway tissues, or place them together in the same cultur...

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Abstract

The invention discloses a method for culturing airway epithelial cells by simply, conveniently and rapidly inducing differentiation of cilia. The method comprises the steps of separating in-vivo airway tissues from mammals, rinsing the airway tissues, longitudinally shearing and separating the airway tissues to form airway rings, tiling the annular airway tissues in a culture vessel, and putting the culture vessel into an incubator of 37 DEG C for culturing. The technical method adopted in the invention is simple, convenient and rapid, the operation procedure is simplified, the culture period is shortened, meanwhile, cultured cells can differentiate a great number of cilia, and a great number of cells can be obtained by virtue of a culture process for researching or other requirements.

Description

technical field [0001] The present invention relates to a method for culturing airway epithelial cells, in particular to a method for separating the airway into independent annular airway tissues by longitudinally cutting the airway for simple and fast culture, and capable of inducing the cultured epithelial cells to differentiate into cilia A method for culturing airway epithelial cells. Background technique [0002] At present, there are two main methods for culturing airway epithelial cells, either by specific enzyme digestion or by cutting and pasting airway tissue. [0003] The main principle of the specific enzyme digestion method is to separate the mammalian airway, use XIV type protease to perfuse the mammalian airway, use the specific digestion effect of XIV type protease on airway epithelial cells, and digest at 37 degrees Celsius for 2 hours or 4 degrees Celsius After overnight digestion for 24 hours, the epithelial cells located on the inner surface of the airwa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0625C12N5/0688
Inventor 李童斐胡清华朱莉萍
Owner 李童斐
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