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Method for removing mycoplasma contamination in tumor cells and application of method

A technology for tumor cells and mycoplasma, which is applied in the field of removing mycoplasma contamination in tumor cells, can solve the problems of short and limited treatment cycle, and achieve the effect of short treatment cycle, simple operation process, and avoidance of antibiotics and high temperature factors.

Pending Publication Date: 2019-10-11
CANCER CENT OF GUANGZHOU MEDICAL UNIV
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Problems solved by technology

[0006] In the current prior art, the strategies for removing mycoplasma contamination in cultured cells are still very limited, and there is no specific method for removing mycoplasma contamination in tumor cells. Therefore, a novel, effective, and less harmful to cells has been developed. It is very meaningful to remove mycoplasma contamination in tumor cells with a short treatment period

Method used

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  • Method for removing mycoplasma contamination in tumor cells and application of method
  • Method for removing mycoplasma contamination in tumor cells and application of method
  • Method for removing mycoplasma contamination in tumor cells and application of method

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Embodiment 1

[0050] This embodiment provides a method for removing mycoplasma contamination in the DU145-GL cell line, specifically comprising the following steps:

[0051] (1) Detect the DU145-GL cell line to be detected with a mycoplasma detection kit, and judge the contamination of the mycoplasma in the cell line. The results are shown in Table 2, B / A>0.9, and the mycoplasma-positive DU145-GL cell line is obtained;

[0052] (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 DU145-GL 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;

[0053] (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 for removing mycoplasma contamination in tumor cells and application of the method. The method comprises the steps that a tumor cell line with the positive mycoplasmas is transplanted into a mouse body with the NSI immune deficiency, the mouse is bred, then the tumor cell line is separated from the inside of the mouse body, and a tumor cell line with the negativemycoplasmas is obtained. Compared with a frequently-used method for removing mycoplasma contamination in cells at present, the provided method is simpler in operation process and safer, the factors ofantibiotics and high temperature are avoided, the damage to the cells can be reduced to the minimum, the treatment period is short, mycoplasma contamination in the tumor cells can also be thoroughlyremoved, and the relapse phenomenon cannot be easily caused.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for removing mycoplasma pollution in tumor cells and an application thereof. Background technique [0002] Mycoplasma is the most common pollutant in cell culture, which is difficult to detect and interfere with experimental results. About 30.3%-50.5% of cell cultures have mycoplasma contamination. Sources of mycoplasma contamination in cultured cells include the use of some products of animal origin such as serum or cell lines that have been contaminated with mycoplasma. The concentration of mycoplasma in the cell culture supernatant can reach 10 7 cells / mL, while the cultured cells look normal, and there is no obvious change in the contaminated cells when observed with the naked eye or under an ordinary light microscope. Due to competition for nutrients and toxic metabolites of mycoplasma, mycoplasma contamination can significantly affect the physiological ac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2509/00
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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