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Method for inducing G-CSFR expression on surface of natural killer cell

A natural killer cell, G-CSFR technology, applied in cell-related fields, can solve the problems of slow natural killer cell proliferation and inability to promote natural killer cell proliferation quickly and efficiently

Pending Publication Date: 2022-02-18
PEOPLES HOSPITAL PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in this scheme, the speed of promoting the proliferation of natural killer cells is relatively slow, and it cannot rapidly and efficiently promote the proliferation of natural killer cells

Method used

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  • Method for inducing G-CSFR expression on surface of natural killer cell
  • Method for inducing G-CSFR expression on surface of natural killer cell
  • Method for inducing G-CSFR expression on surface of natural killer cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Step 1. Flow cytometric detection of the expression of G-CSFR on the surface of NK cells in whole blood

[0049] (1) Take 50-200 μl of fresh heparin anticoagulated blood (depending on the number of cells, and ensure that each tube contains 1×10 6 mononuclear cells) into flow tubes.

[0050] In this step, heparin anticoagulant means that heparin has the effect of preventing blood coagulation.

[0051](2) Add the flow cytometry antibody (CD3 / CD56 / CD114) into the flow cytometry tube in step (1), and incubate at 4° C. for 20 min in the dark. Among them, the role of adding flow cytometry antibodies is to identify cells, specifically: CD3 and CD56 can define NK cells, that is, CD3-CD56+ is NK cells, and CD114 represents the marker of G-CSFR, which can be used to detect NK cells. Expression of G-CSFR.

[0052] (3) Prepare 1×hemolysin, the preparation method is: take 10×hemolysin and sterile water for injection to prepare at a ratio of 1:9, keep it at room temperature, and p...

Embodiment 2

[0107] Step 1. Flow cytometric detection of the expression of G-CSFR on the surface of NK cells in whole blood

[0108] (1) Take 50-200 μl of fresh heparin anticoagulated blood (depending on the number of cells, and ensure that each tube contains 1×10 6 mononuclear cells) into flow tubes.

[0109] In this step, heparin anticoagulant means that heparin has the effect of preventing blood coagulation.

[0110] (2) Add the flow cytometry antibody (CD3 / CD56 / CD114) to the flow cytometry tube in step (1), and incubate at -5°C in the dark for 10 minutes.

[0111] (3) Prepare 1×hemolysin, the preparation method is: take 10×hemolysin and sterile water for injection to prepare at a ratio of 1:9, keep it at room temperature, and prepare freshly before use.

[0112] (4) Add 2ml of 1×hemolysin freshly prepared in step (3) to the flow tube incubated in step (2), mix well, and let it stand for 8 minutes at 20°C to lyse the red blood cells.

[0113] (5) After lysing red blood cells, centrif...

Embodiment 3

[0124] Step 1. Flow cytometric detection of the expression of G-CSFR on the surface of NK cells in whole blood

[0125] (1) Take 50-200 μl of fresh heparin anticoagulated blood (depending on the number of cells, and ensure that each tube contains 1×10 6 mononuclear cells) into flow tubes.

[0126] In this step, heparin anticoagulant means that heparin has the effect of preventing blood coagulation.

[0127] (2) Add the flow cytometry antibody (CD3 / CD56 / CD114) into the flow cytometry tube in step (1), and incubate at 0° C. for 30 minutes in the dark.

[0128] (3) Prepare 1×hemolysin, the preparation method is: take 10×hemolysin and sterile water for injection to prepare at a ratio of 1:9, keep it at room temperature, and prepare freshly before use.

[0129](4) Add 2ml of 1×hemolysin freshly prepared in step (3) to the flow tube incubated in step (2), mix well, and let it stand for 8 minutes at 30°C to lyse the red blood cells.

[0130] (5) After lysing red blood cells, centr...

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Abstract

The invention provides a method for inducing G-CSFR expression on the surface of a natural killer cell, and the method comprises the following steps of: adding an induction factor for inducing G-CSFR expression on the surface of the natural killer cell into a container filled with the natural killer cell, and performing inducing. Compared with the prior art, the method has the advantages that the expression of the G-CSFR on the surface of the natural killer cell is successfully induced through the induction factor for the first time, and the purpose of further promoting the proliferation function of the natural killer cell is achieved by inducing the expression of the G-CSFR on the surface of the natural killer cell.

Description

technical field [0001] The invention belongs to the technical field of cells, and in particular relates to a method for inducing the expression of G-CSFR on the surface of natural killer cells. Background technique [0002] Natural killer cells (NK) are important immune cells in the body, not only related to anti-tumor, anti-viral infection and immune regulation, but also involved in the occurrence of hypersensitivity and autoimmune diseases in some cases. Able to identify target cells and killing mediators. Therefore, how to promote the proliferation of natural killer cells and improve the immune function of the body has attracted the attention of people in the industry. [0003] In the prior art, for the problem of how to promote the proliferation of natural killer cells, the usual technical solution is: cytokines or feeder layer cytotrophy to promote the proliferation of natural killer cells. However, in this scheme, the speed of promoting the proliferation of natural k...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0646C12N2501/2315
Inventor 黄晓军赵翔宇常英军曹勋红
Owner PEOPLES HOSPITAL PEKING UNIV