Cell culture solution and NK cell culture method

A technology of cell culture and culture method, applied in the field of cell culture medium and NK cell culture, can solve the problems of low biological activity and poor expansion effect of NK cells, and achieve the effect of promoting rapid proliferation, simple operation and high biological activity.

Inactive Publication Date: 2017-11-24
GUANGZHOU ZISHENG BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In order to solve the technical problems of poor amplification effect and low biological activity of NK cells in the prior art, the invention provides a method for culturing cell culture fluid and NK cells

Method used

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  • Cell culture solution and NK cell culture method
  • Cell culture solution and NK cell culture method
  • Cell culture solution and NK cell culture method

Examples

Experimental program
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Effect test

Embodiment 1

[0027] 1. Preparation of Peripheral Blood Mononuclear Cells

[0028] (1) Take 30 mL of peripheral blood, after anticoagulant treatment, add normal saline at a ratio of 1:1 between peripheral blood and normal saline, and mix well;

[0029] (2) Take another 50mL centrifuge tube, add 12.5mL of lymphocyte separation medium, and slowly transfer the diluted blood to the surface of the lymphocyte separation medium to form a clear interface between the two (blood dilution: lymphocyte separation medium =2:1), centrifuged at 800g for 15min;

[0030] (3) Take the liquid containing the peripheral blood mononuclear cell (PBMC) layer in the upper layer, be careful not to suck out the red blood cell layer, and transfer the crude peripheral blood mononuclear cell extract to another 50mL centrifuge tube;

[0031] (4) Resuspend the crude extract of peripheral blood mononuclear cells with 40mL RPMI 1640 medium, centrifuge at 500g for 5min, discard the supernatant, then resuspend the peripheral ...

Embodiment 2

[0038] 1. NK cell source and grouping

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Abstract

The invention belongs to the technical field of immune cell culture, and particularly relates to a cell culture solution and an NK cell culture method. The provided cell culture solution includes various kinds of culture solutions including an ALyS505NK-AC culture solution and an ALyS505NK-EX culture solution, wherein the ALyS505NK-AC culture solution containing IL-2 and IL-5 and the ALyS505NK-EX culture solution includes IL-2. The invention further provides the NK cell culture method, the biological activity of NK cells obtained by means of the method is high, the number of amplification is large, therefore, the provided culture method is simple in operation, the survival rate of the NK cells is high, the number of amplification is large, and the culture solution and the culture method can meet various application requirements of clinical or academic research.

Description

technical field [0001] The invention belongs to the technical field of immune cell culture, and in particular relates to a cell culture medium and a culture method for NK cells. Background technique [0002] Natural killer cells (NK cells) are a group of large granular lymphocytes different from T and B lymphocytes, distributed in peripheral lymphoid organs and blood circulation system, and can exert cytotoxic effect without pre-stimulation and activation of antigens , and can secrete a variety of cytokines and chemokines. It is the main bearer of the body's innate immunity and the core regulatory cell of acquired cellular immunity. It plays an important role in tumor immunity, anti-viral infection and removal of non-self cells. NK cell adoptive immunotherapy is an important means of clinical tumor cell immunotherapy. Due to the low content of NK cells in peripheral blood (accounting for about 5% to 10% of lymphocytes), and many factors such as cell concentration, culture m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0646C12N2501/2302C12N2501/2305
Inventor 马颖阮雯莉黄婉娴
Owner GUANGZHOU ZISHENG BIOLOGICAL TECH CO LTD
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