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Method for establishing chicken liver injury cell model

A liver injury and cell model technology, applied in the direction of tumor/cancer cells, artificial cell constructs, animal cells, etc., can solve the problems of complicated separation and culture process, unstable cell state, and restriction of chicken liver damage, so as to reduce economic losses Effect

Pending Publication Date: 2021-12-07
GUANGDONG OCEAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the cells that are often used to construct liver cell injury models are mostly chicken primary hepatocytes. However, chicken primary hepatocytes have disadvantages such as complicated isolation and culture process and unstable cell state in the application process.
The prior art lacks an AFB that is easy to operate and stable in effect 1 Induced Chicken Hepatocyte Injury Model Severely Constrains AFB 1 Molecular Mechanism of Induced Chicken Liver Injury in Vitro and Drug Screening for Prevention and Treatment

Method used

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  • Method for establishing chicken liver injury cell model
  • Method for establishing chicken liver injury cell model
  • Method for establishing chicken liver injury cell model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Establishment of cell model of chicken liver injury:

[0028] (1) The preparation method of the complete culture medium is as follows (50mL system): 44.5mL DMEM / F12 medium + 5mL fetal bovine serum FBS + 0.5mL double antibody (penicillin and streptomycin).

[0029] (2) AFB 1 Dissolved in DMSO and diluted with complete medium to obtain AFB 1 The culture solution with a final concentration of 0.05-0.25 μg / mL and a final concentration of DMSO of 0.01-0.025% is the induction solution.

[0030] (3) Chicken LMH was cultured, and cells with good growth and stable state were obtained for subsequent experiments.

[0031] (4) Take the cells and inoculate them on a cell culture plate, keep the amount of cells in each well consistent, and after the cells adhere to the wall, it occupies about 70-80% of the bottom of the well, add different concentrations of induction solution to the culture wells and cultivate for 6-24 hours.

Embodiment 2

[0033] Cell viability detection:

[0034] The cell survival rate of each group was detected by CCK-8 method, and group 1 was used as the control group, and the other 6 test groups were statistically analyzed. Find the lowest AFB at which cell viability begins to decrease significantly compared to controls 1 The test group where the concentration is and the appropriate action time period. The formula for calculating cell viability is:

[0035]

[0036] The results are shown in the table below:

[0037] Table 1. Effect of LMH on relative cell viability (%)

[0038]

[0039] Note: Different shoulder letters indicate significant difference, P<0.05.

Embodiment 3

[0041] The cell apoptosis rate was detected by flow cytometry, and the results are shown in the table below:

[0042]

[0043] Note: Different shoulder letters indicate significant difference, P<0.05.

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Abstract

The invention discloses a method for establishing a chicken liver injury cell model, which comprises the following steps: 1) preparing a DMEM / F12 culture medium containing fetal calf serum with the final concentration of 10%, the culture medium containing penicillin and streptomycin; 2) dissolving aflatoxin B1 in a dimethyl sulfoxide solution and diluting the aflatoxin B1 in a dimethyl sulfoxide solution with the culture medium in the step 1), obtaining an induction solution, wherein the final concentration of aflatoxin B1 in the induction solution is 0.05-0.25 microgram / mL, and the final concentration of dimethyl sulfoxide in the induction solution is 0.01-0.025%; 3) culturing the chicken liver cancer cell line until the growth vigor is good and the state is stable; and 4) when the chicken hepatoma carcinoma cell line grows to 70-80% of the adherence, adding the induction liquid in the step 2), and performing induction culture for 6-24 hours to obtain the chicken hepatoma carcinoma cell line.

Description

technical field [0001] The invention belongs to the field of cell model establishment, in particular to a kind of aflatoxin B 1 (AFB 1 )-induced cell model of chicken liver injury. Background technique [0002] According to the investigation of the Food and Agriculture Organization of the United Nations (FAO), more than 25% of the grains in the world are polluted by mycotoxins (Mycotoxins), among which aflatoxin (Aflatoxin, AF) is the most serious. According to the investigation and research on feed raw materials in my country, the AF pollution ratio in corn in China is as high as 84%. AF is a secondary metabolite of Aspergillus flavus and Aspergillus parasiticus, about 20 species of AF have been found, among them, AFB 1 It is listed as a first-class carcinogen because of its strongest toxicity, greatest harm and widest distribution. At the same time, AFB 1 It is also the most potential cytotoxic substance. It enters the body to generate reactive oxygen species (ROS) t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12N5/071
CPCC12N5/0693C12N5/067C12N2500/62C12N2501/999
Inventor 刘文超郭艳赵志辉高振赵越邱盛坚金永燕
Owner GUANGDONG OCEAN UNIVERSITY
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