Method for detecting influence of electronic cigarette product on micronucleus rate of cells

A technology of electronic cigarettes and micronucleus rate, applied in the field of biotechnology applications, can solve the problems of not meeting the needs of genotoxicity testing of electronic cigarette products, and not formulating standard methods for in vitro micronucleus testing of electronic cigarette products

Active Publication Date: 2017-03-29
CHINA TOBACCO YUNNAN IND
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AI Technical Summary

Problems solved by technology

No smoke is produced during use, and its use method is significantly different from traditional cigarettes. Changes in the test object lead to differences in sample pretreatment methods, test cells, and test doses. In vitro micronucleus test for the detection of genotoxic

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  • Method for detecting influence of electronic cigarette product on micronucleus rate of cells

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Embodiment Construction

[0044] In this example, the micronuclei of human lung fibroblasts were tested for four foreign commercially available e-cigarette liquids as follows:

[0045] (1) Pretreatment of the test substance: Take a certain amount of electronic cigarette stock solution, mix and dilute it according to the ratio of the volume ratio of electronic cigarette stock solution and serum-free cell culture medium to 70%, leave it for 32 hours, and filter it out with a sterile filter. Bacteria, the solution to be tested was obtained, and stored at -80°C until use;

[0046] (2) Preparation of single-cell suspension of human lung fibroblasts: After resuscitating human lung fibroblasts, inoculate them into cell culture flasks and place them at 37°C and 5vt% CO 2 Cultivate in an incubator, observe the confluence and morphology of the cultured cells with an inverted microscope, remove the medium in the culture flask when the cells grow to 90% confluence, wash twice with phosphate buffer, add an appropri...

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Abstract

The invention discloses a method for detecting the influence of an electronic cigarette product on the micronucleus rate of cells. The method comprises the following steps: preprocessing test samples, preparing a human lung fibroblast single cell suspension fluid, calculating the concentration of the human lung fibroblast single cell suspension fluid, inoculating the human lung fibroblast single cell suspension fluid, dividing the test samples into groups, setting the detection dosages of the test samples, preparing a test sample culture medium, subjecting the test sample culture medium to low permeability, and preparing a droplet slice of the test sample culture medium. The using characteristics and effect modes of electronic cigarette products are taken into account, an electronic cigarette product sample preprocessing method is established; according to the electronic cigarette product acted target cells, human lung fibroblast is taken as the cell for the cytotoxicity tests of electronic cigarette products, the sample detection dosage is determined at the same time, and thus an in-vitro micronucleus test method suitable for the electronic cigarette products is established.

Description

technical field [0001] The invention belongs to the application field of biotechnology, and in particular relates to a method for detecting the influence of electronic cigarette products on the micronucleus rate of cells. Background technique [0002] Micronucleus test has a clear detection end point, simple method, and easy to carry out. It is one of the standard combination tests of genotoxicity. Damage detection and early forecasting have been widely used. In 2009, the Organization for Economic Co-operation and Development (OECD) published a guidance document for in vitro micronucleus tests in mammals, initially establishing a unified standard for in vitro micronucleus tests in mammals. In 2002, CORESTA, the International Tobacco Scientific Research Cooperation Center, established a working group for in vitro toxicological testing of cigarette smoke. After a large number of literature research and research work, the working group recommended the use of in vitro micronucl...

Claims

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Application Information

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IPC IPC(8): G01N33/50
CPCG01N33/5014
Inventor 李雪梅管莹夭建华熊国行米其利高茜朱洲海
Owner CHINA TOBACCO YUNNAN IND
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