Tumor cell proliferation detection method based on Hochest333258 and application

Abstract

The invention discloses a tumor cell proliferation detection method based on Hochest333258 and application. The method comprises the following steps: subculturing tumor cells to a cell culture plate,counting cells, then carrying out doubling dilution, so that a gradient series with different cell numbers is obtained, carrying out the conventional culture in a culture medium, then adding liquid containing Hochest333258, uniformly mixing, then standing for a period of time, abandoning a cell culture medium, washing off residual culture medium with PBS buffer liquid, then adding the PBS buffer liquid, finally detecting a fluorescent value, and judging cell proliferation level or survival rate by virtue of fluorescence signal intensity. The method disclosed by the invention is simple and is high in repeatability and accuracy; and the proliferation level of the tumor cells can be effectively detected.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for detecting cell proliferation based on Hochest333258-labeled tumor cells and its application. Background technique [0002] The level of tumor proliferation is closely related to the degree of malignancy of tumor cells, and it is an important detection index for anti-tumor research and treatment. In the screening and identification of anti-tumor drugs and the research of tumor therapeutic targets, the detection of tumor cell proliferation level is a basic research method. At present, the commonly used method is: use the tumor cell line cultured in vitro as a model, and use MTT method, CCK-8 method, cell counting method or crystal violet staining method to detect after a period of intervention with drug treatment, gene regulation and other conditions, The number of viable tumor cells. Then, the survival rate or inhibition rate of the tumor cells is calculated by comparing ...

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Problems solved by technology

But its disadvantage is that certain treatment conditions may lead to changes in the activity of enzymes in the cells, which directly affects the experimental results

In addition, this method has many processing steps, which easily leads to increased experimental errors.

The standard deviation of the multiple well data of CCK-8 and MTT experiments is usually large, which may easily lead to no statistical difference in the data due to errors, so the requirements for the experimental operation are relatively high

[0006] The advantage of cell counting method is intuitive and simple, but it cannot perform high-throughput multi-sample operation, and is not suitable for large-sample control analysis such as drug screening, so its application is extremely limited

In addition, due to the need to digest the cells into a single cell suspension, the experimental error is large

[0007] The advantage of crystal violet staining is that it is the most convenient, but it cannot distinguish between living cells and dead cells, and is interfered by residual crystal violet staining solution and staining efficiency, and the experimental error is also large.

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