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Method capable of improving cell state and application thereof

A cell state and cell technology, applied in the biological field, can solve the problem of limited methods for removing mycoplasma contamination, and achieve the effect of removing mycoplasma contamination and simple operation process

Pending Publication Date: 2019-09-24
CANCER CENT OF GUANGZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] It can be seen that the method for removing mycoplasma pollution in the prior art is still very limited, and there are many disadvantages. Therefore, a novel, safer, less harmful to cells, short treatment cycle, and can be completely removed The method of mycoplasma contamination is very meaningful

Method used

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  • Method capable of improving cell state and application thereof
  • Method capable of improving cell state and application thereof
  • Method capable of improving cell state and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0061] The present embodiment provides a method for removing mycoplasma contamination in the MDA-MB-231-GL cell line, specifically comprising the following steps:

[0062] (1) Detect the MDA-MB-231-GL cell line to be detected with a mycoplasma detection kit, and judge the contamination situation of mycoplasma in the cell line. 231-GL cell line;

[0063] (2) Moisten and disinfect the skin at the groin of the hind leg of the NSI immunodeficient mouse after dipping it in alcohol with an alcohol cotton ball, draw 200 μL of the mycoplasma-positive MDA-MB-231-GL cell line in step (1) with an insulin needle, Take 2×10 5 The amount of cells injected into the mouse subcutaneously at an angle of 45 degrees, a bulging mass can be seen, and the mouse is put back into a normal environment and raised for 20 days;

[0064] (3) The mice reared in step (2) were killed by dislocation of the cervical vertebrae and the surface was sterilized. In the biological safety cabinet, the skin around th...

Embodiment 2

[0072] The present embodiment provides a method for removing mycoplasma contamination in the HepG2 cell line, specifically comprising the following steps:

[0073] (1) Detect the HepG2 cell line to be detected with a mycoplasma detection kit, and judge the contamination situation of mycoplasma in the cell line. The results are as shown in Table 6, B / A>0.9, and the Mycoplasma-positive HepG2 cell line is obtained;

[0074] (2) Moisten and disinfect the skin at the groin of the hind legs of NSI immunodeficient mice with alcohol cotton balls dipped in alcohol, draw 200 μL of the mycoplasma-positive HepG2 cell line in step (1) with an insulin needle, and dilute 2×10 5 The amount of cells injected into the mouse subcutaneously at an angle of 45 degrees, a bulging mass can be seen, and the mice are put back into the normal environment and raised for 40 days;

[0075] (3) The mice reared in step (2) were killed by dislocation of the cervical vertebrae and the surface was sterilized. I...

Embodiment 3

[0082] The present embodiment provides a method for removing mycoplasma contamination in the Huh7 cell line, specifically comprising the following steps:

[0083] (1) Detect the Huh7 cell line to be detected with a mycoplasma detection kit, and judge the contamination situation of mycoplasma in the cell line. The results are as shown in Table 6, B / A>0.9, and the mycoplasma-positive Huh7 cell line is obtained;

[0084] (2) Moisten and disinfect the skin at the groin of the hind legs of NSI immunodeficient mice with alcohol cotton balls dipped in alcohol, draw 200 μL of the mycoplasma-positive Huh7 cell line in step (1) with an insulin needle, and dilute 2×10 6 The amount of cells injected into the mouse subcutaneously at an angle of 45 degrees, a bulging mass can be seen, and the mice are put back into the normal environment and raised for 30 days;

[0085] (3) The mice reared in step (2) were killed by dislocation of the cervical vertebrae and the surface was sterilized. In th...

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Abstract

The invention relates to a method capable of improving the cell state and application thereof. The method comprises the steps of transplanting a mycoplasma positive cell line into the body of an immunodeficient mouse, raising the mouse, and separating a cell line from the body of the mouse to obtain a mycoplasma negative cell line. Compared with current common methods for removing mycoplasma contamination in cells, the method for removing mycoplasma contamination in cells has a simpler and safer operation flow; meanwhile, the method avoids antibiotic and high-temperature factors, can minimize the damage to cells, has a shorter treatment cycle and can also completely remove mycoplasma contamination without a relapse.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for improving cell state and its application. Background technique [0002] Cell culture is a routine experimental technique for expanding cells in vitro, and is currently widely used in scientific research and clinical experiments. One of the keys to the success of cell culture is to control pollution. Bacteria, mold, black colloid, mycoplasma, etc. are all microorganisms that cause contamination of cultured cells, and mycoplasma pollution is a major source of pollution in the process of cell culture. [0003] Mycoplasma is a class of minimal prokaryotic microorganisms without cell walls, highly pleomorphic, able to pass through bacteria filters, and can be cultured and proliferated in artificial medium. Because it can form filamentous and branched shapes, it is called mycoplasma. Mycoplasma widely exists in humans and animals, and most of them are not pathoge...

Claims

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Application Information

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IPC IPC(8): C12N5/0781C12N5/0783C12N5/09
CPCC12N5/0693C12N5/0635C12N5/0636C12N5/0646C12N2509/00
Inventor 李鹏黄晓涵蒋治武姚瑶程琳王素娜吴绮婷龙有国黄启键林思妙
Owner CANCER CENT OF GUANGZHOU MEDICAL UNIV
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