Method for detecting instability of human cell mitotic chromosomes in vitro

An in vitro detection and mitosis technology, applied in the field of life sciences, can solve the problems of reduced sensitivity, high economic cost, labor-intensive, etc., and achieve the effect of reducing false positives, low economic cost, and labor-saving

Inactive Publication Date: 2017-05-24
YUNNAN NORMAL UNIV
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Problems solved by technology

Although this method can analyze and count most of the chromosomal instability phenomena, the disadvantages of this method are: ① labor-intensive
The chromosome dispersion method has high requirements on the state of chromosome separation, and the number of chromosomes in human cells is large and the size varies greatly. These factors limit the num

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  • Method for detecting instability of human cell mitotic chromosomes in vitro
  • Method for detecting instability of human cell mitotic chromosomes in vitro

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Embodiment Construction

[0031] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some, not all, embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without creative efforts fall within the protection scope of the present invention.

[0032] A method for in vitro detection of mitotic chromosome instability in human cells, the steps comprising:

[0033] 1. Cell processing process

[0034] (1) Cell culture: NCM460 cells use RPMI-1640 medium, add 10% newborn bovine serum, 1% glutamine of 2mM and 0.1% penicillin and 5mg / mL streptomycin of 5000IU / mL, NCM460 cells Cultivated at 75cm 2 In a glass or polystyrene culture flask, place CO 2 In a humid incubator with a content of 5% and a temperature of ...

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Abstract

The invention provides a method for detecting the instability of human cell mitotic chromosomes in vitro. The method comprises the following steps: culturing cells, treating the cells, collecting the cells, concentrating the cells, smearing the cells, fixing the cells, dyeing the cells and sealing the cells; observing mitotic cells by utilizing an optical microscope and carrying out frequency counting on an instability index of the chromosomes. The method is a method for systemically and omnidirectionally detecting the instability of the mitotic chromosomes; compared with other methods, the method has the advantages that (1) an instability phenomenon of a plurality of types of chromosomes can be detected at the same time, and whether a certain compound is a chromosome toxin and/or a mitotic toxin is analyzed; (2) the method has very high sensitivity and can be used for detecting extremely low dosage of genetic toxic substances; (3) a false positive phenomenon caused by artificial reasons can be extremely reduced; (4) the method has the characteristics of simplicity and easiness of operation; the labor force can be effectively saved and the method has a high-throughput application potential; (5) the method has relatively low economic cost and is applicable to development and implementation in most of laboratories.

Description

technical field [0001] The invention relates to the technical field of life sciences, in particular to a method for detecting chromosome instability in human cell mitosis in vitro. Background technique [0002] Mitosis is the most vulnerable phase of the cell cycle as cells are exposed to exogenous or endogenous chemicals that cause chromosomal instability. Chromosomal instability can further lead to genomic instability in cells, which is the underlying cause of many human genetic-environmental diseases, which are associated with birth defects, developmental abnormalities, immune deficiencies and degenerative diseases (such as Alzheimer's disease, Parkinson's disease , hypertension, cardiovascular disease, diabetes, osteoporosis, tumors, etc.) are highly correlated with increased risk. Because there are many indicators of chromosomal instability, most of the detection methods currently used can only target a few of them, and there is a lack of methods that can systematicall...

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q1/6886C12Q2600/142
Inventor 倪娟郭锡汉汪旭薛京伦
Owner YUNNAN NORMAL UNIV
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