Cigarette smoke in-vitro micronucleus detection method

A technology for micronucleus detection and cigarette smoke, which is applied to the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of difficult control of the concentration of the test substance, complicated test process, inconsistent exposure methods and routes, etc. The effect of shortening the test time, enriching the experimental results, and reducing the experimental cost

Inactive Publication Date: 2010-03-17
ZHENGZHOU TOBACCO RES INST OF CNTC
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Problems solved by technology

[0007] The applicant in this case also applied for a patent titled "In vivo Micronucleus Analysis and Detection Method in Mice for Evaluating Cigarette Smoke Toxicity", the application number of which is 200810049452.3. To investigate the change of chromosome integrity and chromosomal segregation of cigarette smoke by way of gastrointestinal exposure, the way of exposure is inconsistent with the actual route of exposure to cigarette smoke, the test process is complicated, and the concentration of the test substance is not easy to control

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  • Cigarette smoke in-vitro micronucleus detection method

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[0024] The reference cigarette 2R4F was selected for in vitro micronucleus test, and CHO was cultivated until the concentration reached 1×10 5 cells / mL, the cell suspension was inoculated in a 50 mL culture flask at 5 mL / bottle. Cells were incubated overnight to allow attachment.

[0025] After the cigarette smoke condensate was extracted with DMSO, the extract was diluted with cell culture medium to prepare different doses (50, 75, 100, 150, 200 μg / mL) of the cigarette smoke condensate. After the cells grow on the wall, discard the original culture medium, add the test substance, inoculate two culture flasks for each dose, and set up a negative control and a positive control. When activation is required, add 4.625 medium and 0.275mL S9 mixture into the culture bottle. Cultivate for 27h.

[0026] After the culture, the cells were digested, smeared on a glass slide, and the cells were fixed with a fixative. Then stain with acridine orange stain for 5min, rinse the stain, an...

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Abstract

The invention relates to a cigarette smoke in-vitro micronucleus detection method. The cigarette smoke in-vitro micronucleus detection method is characterized in that Chinese hamster ovary cells are selected to carry out the micronucleus test, and the micronucleus frequency is detected by cell culture, preparation of cigarette smoke condensate, cell contamination, fixing and dyeing. Compared withthe prior art, the cigarette smoke in-vitro micronucleus detection method has the advantages that in-vitro micronucleus test is adopted to evaluate the influence of cigarette smoke on the cell chromosomal aberration, not only the experiment cost can be reduced, the test time can be shortened and richer experiment result can be obtained, but also the in-vitro micronucleus test can obtain the resultconsistent to the in-vivo micronucleus test, as well as equivalent sensibility, specificity and accuracy; besides, the source of the micronucleus can be analyzed on basis of the obtaining of the cellmicronucleus frequency.

Description

technical field [0001] The invention relates to a cigarette toxicity detection method, in particular to an in vitro micronucleus test method for cigarette smoke condensate. Background technique [0002] Micronucleus is a micronucleus after the chromosome of the cell breaks, and when the cell enters the next division, the chromosome fragment cannot enter the daughter cell with mitosis, but forms a micronucleus in the cytoplasm whose diameter is less than 1 / 3 of the main nucleus. Round or oval tiny nuclei separate from the main nucleus. Micronucleus determination is a simple and rapid cytogenetic examination method, which is important in the diagnosis and prevention of liver cancer, esophageal cancer, lung cancer and other malignant tumors, as well as the evaluation of cytotoxic substances such as drugs, radiation and smoking on human, animal and in vitro cell damage. significance. [0003] In 1923, Taylor C.V. discovered the micronucleus for the first time and explained the...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 尚平平李翔聂聪彭斌赵乐刘惠民谢剑平
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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