Method for establishing in-vitro detection model of chemical-induced human esophageal cancer

A technology for in vitro detection and establishment of methods, which is applied in biochemical equipment and methods, measurement/testing of microorganisms, microorganisms, etc., can solve the problems of lack of efficient and accurate in vitro experimental methods, difficulties in observing complex effect end points, etc., and achieve the detection process Efficient and accurate, satisfying high-throughput detection, and the effect of cumbersome detection process

Inactive Publication Date: 2019-02-01
CHINA NAT CENT FOR FOOD SAFETY RISK ASSESSMENT
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Current in vitro alternatives for toxicology testing are used in acute toxicity testing such as skin irritation testing, but have difficulty in chronic toxicity and carcinogenicity testing or in observing complex effect endpoints
Therefore, there is a lack of efficient and accurate in vitro experimental methods to clarify the relationship between chemicals and the induction of human esophageal cancer

Method used

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  • Method for establishing in-vitro detection model of chemical-induced human esophageal cancer
  • Method for establishing in-vitro detection model of chemical-induced human esophageal cancer
  • Method for establishing in-vitro detection model of chemical-induced human esophageal cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0064] (1) Cell culture of human esophageal epithelial cells

[0065] Human normal esophageal cell line Het-1A (ATCC) was cultured in Bronchial epithelial cell basalmedium (BEGM) medium. Culture vessel: 25cm 2 Culture flask, use after coating with 0.1% gelatin. Culture conditions: 95% air, 5% carbon dioxide, 37C. During the cell culture period, the culture medium was changed every 3 days, and the cells were subcultured when the cells grew to cover 80% of the area of ​​the culture bottle. When the cells are subcultured, wash the cells twice with PBS, digest with trypsin until the cells become round, collect the cells after the BEGM medium terminates the reaction, centrifuge at 500g for 5 minutes, discard the supernatant, and add new culture medium for subculture at a ratio of 1:3. . When the cells are cryopreserved, select the cells with good growth status and vigorous growth, wash the cells twice with PBS, digest with trypsin until the cells become round, stop the reaction...

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Abstract

The invention discloses a method for establishing an in-vitro detection model of chemical-induced human esophageal cancer. The method comprises the steps of: providing human esophageal epithelial cells inoculated in an in-vitro culture container; respectively applying the to-be-tested chemicals with a plurality of concentrations in a predetermined concentration range to the human esophageal epithelial cells for culturing for a predetermined time, and measuring a change curve of the cell viability of the human esophageal epithelial cells followed as the concentration of the to-be-tested chemicals; selecting a concentration interval of the to-be-tested chemicals corresponding to a predetermined cell viability range in the curve as a carcinogenic determination concentration interval, and selecting a plurality of concentrations in the carcinogenic measurement concentration interval as the carcinogenic assay concentration; respectively applying the to-be-tested chemicals with the carcinogenic assay concentration to the human esophageal epithelial cells to test the carcinogenicity test of the human esophageal epithelial cells by the to-be-tested chemicals.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for establishing an in vitro detection model of human esophagus cancer induced by chemicals. Background technique [0002] Cancer is usually the result of the interaction between genetic factors and environmental factors. It is generally believed that 90% of human cancers are related to environmental factors, among which chemical factors account for more than 90%. At present, the long-term carcinogenicity test in rodents is the only standard evaluation method for the carcinogenicity of chemicals accepted by regulatory agencies in various countries. Although other test methods such as structure-activity relationship analysis, genotoxicity test, in vitro cell transformation test and population tumor epidemiological investigation are used to evaluate the carcinogenicity of chemicals, there is no ideal test system that can replace rodent carcinogenicity. test. Chemicals are th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02C12N5/09
CPCC12N5/0693C12N2500/02G01N33/5017
Inventor 杨辉贾旭东张倩男
Owner CHINA NAT CENT FOR FOOD SAFETY RISK ASSESSMENT
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