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Method for evaluating cell killing efficacy based on cell diameter and application thereof

A cell and potency technology, applied in the field of cell killing potency detection, can solve the problems of inability to collect cell images and reduce the repeatability of experimental results, and achieve the effects of intuitive and accurate results, simple operation and high application value.

Inactive Publication Date: 2021-01-29
SHANGHAI RUIYU BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, flow cytometer is a liquid flow system and cannot collect cell images. If you need to verify the accuracy of the results, you need to use a microscope or other instruments to confirm the results.
At the same time, in the process of detecting cell killing, there are many subjective errors in the method of flow cytometry; for example, in the experimental stage, the operator needs to adjust the voltage according to experience; Subjective errors lead to reduced repeatability of experimental results

Method used

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  • Method for evaluating cell killing efficacy based on cell diameter and application thereof
  • Method for evaluating cell killing efficacy based on cell diameter and application thereof
  • Method for evaluating cell killing efficacy based on cell diameter and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] This example provides a method for evaluating cell killing efficacy based on cell diameter.

[0065] Described method specifically comprises the steps:

[0066] (1) Prepare effector cells as effector cell suspension, the maximum diameter of the effector cells is 9 μm;

[0067] (2) Collect tumor cells, prepare target cell suspension, the minimum diameter of the target cells is 9 μm, and transfer to 96-well plate;

[0068] (3) Set the effector-target ratio to 1:1, and add the effector cell suspension into the 96-well plate that already has the target cell suspension;

[0069] (4) After 24 hours of co-cultivation, remove the supernatant, wash with PBS buffer, add 0.25% trypsin to digest the adherent cells, centrifuge, resuspend, and adjust the cell density to 1×10 6 / ml.;

[0070] (5) After mixing the cell suspension with 0.2% trypan blue solution at a volume ratio of 1:1, pipette 20 μL to the counting plate and place it in the detection instrument (the instrument used ...

Embodiment 2

[0084] This example provides a method for evaluating cell killing efficacy based on cell diameter.

[0085] Described method specifically comprises the steps:

[0086] (1) Prepare effector cells as effector cell suspension, the maximum diameter of the effector cells is 9 μm;

[0087] (2) Collect tumor cells, prepare target cell suspension, the minimum diameter of the target cells is 9 μm, and transfer to 96-well plate;

[0088] (3) Set the effector-target ratio to 1:1, and add the effector cell suspension into the 96-well plate that already has the target cell suspension;

[0089] Prepare the control group cells at the same time: add only the target cells and the medium to the culture dish as the target cell control group, and add only the effector cells and the medium to another culture dish as the effector cell control group;

[0090] The above-mentioned control group and experimental group are placed under the cultivation environment and cultivated;

[0091] (4) After 24...

Embodiment 3

[0112] Compared with Example 2, in this example, different effect-to-target ratios (E:T) were set as experimental groups to measure the killing efficacy of cells under different effect-to-target ratios.

[0113] The working concentration of effector cells was adjusted, and the effect-to-target ratio was set to 10:1, 5:1 and 1:1, and the remaining steps and experimental conditions were kept the same as in Example 2.

[0114] The obtained experimental results can determine the best effect-to-target ratio and provide a basis for further screening or optimization. To sum up, the method provided by the present invention is relatively intuitive, and can directly obtain visualized cell fluorescence images, obtain different states of cells based on microscopic images, and then analyze the killing efficacy of immune cells. The method has simple steps and improves detection efficiency and accuracy.

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Abstract

The invention provides a method for evaluating cell killing efficacy based on cell diameter and application thereof. The method comprises the following steps: mixing and co-culturing target cells andeffector cells to obtain a cell suspension, and adding a cell dye to dye dead cells in a cell suspension; carrying out microscopic imaging on the dyed cells to obtain a microscopic image; and recognizing cells in the microscopic image through image recognition, collecting size information and color information of the cells, analyzing the obtained information, and evaluating the cell killing efficacy of effector cells according to an analysis result. The method provided by the invention combines bright field microscopic imaging and image analysis technologies, can directly analyze microscopic images and obtain detection results, the obtained results are more accurate and intuitive, and the method has important application value in the fields of medical diagnosis and biomedicine industrialization which need standardization and repeated verification.

Description

technical field [0001] The invention relates to the technical field of cell killing efficacy detection, in particular to a method for evaluating cell killing efficacy based on cell diameter and an application thereof. Background technique [0002] The evaluation of cell killing efficacy, that is, the detection of cytotoxicity, is widely used in biological, clinical and pharmaceutical research. In immunotherapy, it is an essential step to determine the cell viability and activity of immune effector cells by evaluating the cell killing efficacy. [0003] At present, there are many evaluation methods for cell killing efficacy, including: MTT reduction method, lactate dehydrogenase (LDH) release method, reporter gene transfection method, ELISPOT test and radioactive chromium (51Cr) release assay, etc. Among them, the most popular assay for assessing cell killing potency is the 51Cr release assay, which is considered the “gold standard” for detecting cell-mediated cytotoxicity, ...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6402G01N21/6486
Inventor 姜晶罗浦文陈凯
Owner SHANGHAI RUIYU BIOTECH
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