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
View PDF5 Cites 2 Cited by
  • 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

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • 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

Example Embodiment

[0063]Example 1

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

[0065]The method specifically includes the following steps:

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

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

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

[0069](4) After 24 hours of co-cultivation, remove the supernatant and wash with PBS buffer, add 0.25% trypsin to digest the adherent cells, centrifuge, resuspend, and adjust the cell density to 1×106 / ml.;

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

Example Embodiment

[0083]Example 2

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

[0085]The method specifically includes the following steps:

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

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

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

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

[0090]Place the above-mentioned control group and experimental group in a culture environment;

[0091](4) After 24 hours of co-cultivation, remove the supernatant and w...

Example Embodiment

[0111]Example 3

[0112]Compared with Example 2, in this example, different effective-to-target ratios (E:T) are also set as the experimental group to determine the killing efficacy of cells under different effective-to-target ratios.

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

[0114]The experimental results obtained can determine the best effective target ratio and provide a basis for subsequent further screening or optimization. In summary, 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 the microscopic images, and then analyze the killing efficacy of immune cells. The method has simple steps and improves the efficiency and accuracy of detection.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
The maximum diameteraaaaaaaaaa
Login to view more

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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N21/64
CPCG01N21/6402G01N21/6486
Inventor 姜晶罗浦文陈凯
Owner SHANGHAI RUIYU BIOTECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap