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Method for detecting killing effect of gamma-delta T cells to tumor cells by utilizing real-time cell analysis system

A tumor cell and analysis system technology, which is applied in the field of detecting the killing effect of γδT cells on tumor cells, can solve the problems of misjudgment of results and differences in operation methods, and achieve the effect of good stability and high sensitivity

Inactive Publication Date: 2016-01-20
杭州庆正鸿科技有限公司
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Problems solved by technology

At present, the commonly used methods for evaluating the cytotoxic effect of γδT cells include MTT method, LDH method, time-resolved fluorescent immunoassay, etc., but these methods do not carry out real-time and continuous monitoring, so the difference in cell state and operation method may be different. cause errors, leading to misjudgment of the results

Method used

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  • Method for detecting killing effect of gamma-delta T cells to tumor cells by utilizing real-time cell analysis system
  • Method for detecting killing effect of gamma-delta T cells to tumor cells by utilizing real-time cell analysis system

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Embodiment 1

[0020] 1) At 37°C, 5% CO 2 Culture A549 cells in the environment to logarithmic growth phase;

[0021] 2) Add 95 μl of 1640 culture medium to each well of the E-plate detection plate, incubate the E-plate detection plate at 37°C for 0.5 hours, and measure the background impedance value;

[0022] 3) Dilute an appropriate amount of logarithmic phase A549 cells to 50,000 cells / ml, inoculate 100 μl of cell suspension in each well, and monitor cell growth in real time;

[0023] 4) After 24 hours, according to the cell growth index, it can be judged whether the cells have entered the logarithmic phase. When the cells have entered the logarithmic phase, select plate wells with consistent cell growth conditions and add 5 μl of different concentrations of γδT cells to each well according to different effect-to-target ratios. ;

[0024] 5) Continue real-time monitoring, and terminate the experiment after the cells in the blank control group enter the apoptotic phase.

[0025] The kil...

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Abstract

The invention discloses a method for detecting the killing effect of gamma-delta T cells to tumor cells by utilizing a real-time cell analysis system and belongs to the technical field of biological medicines. The method is characterized by using the Real-Time Cell Analysis system (RTCA) to detect the killing effect of the gamma-delta T cells to the tumor cells. The method disclosed by the invention has the advantages of high sensitivity, good stability and capability of detecting the killing effect of the gamma-delta T cells to the tumor cells continuously and dynamically in real time without marks.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for detecting the killing effect of γδ T cells on tumor cells by using a real-time cell analysis system. Background technique [0002] γδT cells are a subset of T lymphocytes discovered in recent years, accounting for about 1% to 10% of the total number of T lymphocytes in normal human peripheral blood. There are two main subgroups of human γδT cells, namely Vδ1T and Vδ2T cells . Vδ1T cells are mainly distributed in human epithelial tissue, and Vδ2T cells are mainly distributed in human peripheral blood. Vγ9Vδ2 T cells accounted for more than 90% of peripheral blood γδT cells. With the deepening of research on the structure and function of γδT cells, it has been found that γδT cells can directly recognize protein and peptide antigens and non-peptide antigens without the need for MHC molecule presentation, and have some characteristics of antigen-present...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02
Inventor 吴艳玲张文丁燕萍程行健申立文
Owner 杭州庆正鸿科技有限公司
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