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Laying hen oviduct ampulla epithelial cell culture and oxidative stress model establishment method

An epithelial cell and culture method technology, applied in cell culture active agents, artificial cell constructs, biochemical equipment and methods, etc., can solve the problems of egg white quality decline, fibroblast contamination, etc. Reasonable and effective overcoming susceptibility to fibroblast contamination

Active Publication Date: 2016-11-30
SICHUAN AGRI UNIV
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Problems solved by technology

[0005] In view of this, the present invention aims at the problem that the epithelial cells of the enlarged oviduct of laying hens cultured by the existing method are easily polluted by fibroblasts, and in order to further explore the specific mechanism of the decline in the quality of egg white caused by the external environment, a kind of enlarged oviduct of laying hens is provided. Method for culture of epithelial cells and establishment of oxidative stress model

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  • Laying hen oviduct ampulla epithelial cell culture and oxidative stress model establishment method
  • Laying hen oviduct ampulla epithelial cell culture and oxidative stress model establishment method
  • Laying hen oviduct ampulla epithelial cell culture and oxidative stress model establishment method

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Embodiment Construction

[0045] The implementation of the present invention will be described in detail below with examples, so as to fully understand and implement the implementation process of how the present invention uses technical means to solve technical problems and achieve technical effects.

[0046] The method for cultivating epithelial cells of the enlarged oviduct of laying hens of the present invention adopts Roman laying hens as experimental animals, and specifically comprises the following steps:

[0047] Step 1, after the laying hen is put to death, the entire oviduct is taken out under aseptic conditions and soaked in PBS, and then the mesentery, connective tissue and blood are removed in a biological safety cabinet;

[0048] Step 2: Cut off the swollen part of the fallopian tube, wash it twice inside and outside with phosphate buffered saline (PBS) (Hyclone) preheated at 37°C, and then use ophthalmic scissors to cut the swollen part to 1mm 3 Left and right tissue blocks, during which ...

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Abstract

The invention discloses a laying hen oviduct ampulla epithelial cell culture and oxidative stress model establishment method. The cell culture method consists of: soaking a whole section of laying hen oviduct in PBS, and removing mesentery, connective tissue and blood; cutting off the oviduct ampulla, performing cleaning and then cutting it into tissue pieces; transferring the cleaned tissue pieces into collagenase IV for digestion; filtering the digestive juice, centrifuging the filtrate and discarding the supernate, adding a complete medium to resuspend the cell, and finally transferring the cell into a culture bottle to conduct cultivation. The oxidative stress model establishment method consists of: testing the heavy metal of different concentrations and the cell viability under different action time, and determining the action time point; designing different heavy metal concentration gradients according to the time point, determining cell apoptosis and other conditions, and establishing an oxidative stress model. The invention establishes the laying hen oviduct ampulla epithelial cell oxidative stress model for further exploration of the specific mechanism for egg white quality decline caused by the external environment.

Description

technical field [0001] The invention belongs to the technical field of cytotoxicity, and in particular relates to a method for culturing epithelial cells of the enlarged oviduct of laying hens, and also relates to a method for establishing an oxidative stress model of epithelial cells of the enlarged oviduct of laying hens. Background technique [0002] Eggs are the main source of protein for humans. Changes in egg quality may be accompanied by egg safety issues. With the improvement of living standards, people pay more and more attention to the quality and safety of eggs, and the rapid development of modern industry has aggravated the pollution of the environment, and the safety of common human foods such as eggs is threatened. This not only has a fatal blow to the laying hen industry and even the entire animal husbandry industry, but also has a certain impact on human health. Eggs are formed in the fallopian tube except for the yolk. The fallopian tubes of laying hens a...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071G01N33/50
CPCC12N5/0625C12N2500/32C12N2500/80C12N2501/11C12N2501/33C12N2501/91G01N33/5008G01N33/5044G01N2510/00G01N2800/7009
Inventor 黄选洋王建萍张克英丁雪梅白世平曾秋凤
Owner SICHUAN AGRI UNIV
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