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Serum-free culture medium of NK (Natural Killer) cells

A serum-free medium and NK cell technology, which is applied in the direction of cell culture active agent, culture process, tissue culture, etc., can solve the problems of unstable medium components, achieve good functional consistency, stable components, and increase the value-added multiple Effect

Inactive Publication Date: 2018-02-13
英普乐孚生物技术(上海)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In view of the above-mentioned and / or other problems in the related art, the present invention provides a serum-free medium for NK cells with more stable components and a preparation method thereof, which improves the instability of the medium components caused by the complex composition of albumin Case

Method used

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  • Serum-free culture medium of NK (Natural Killer) cells
  • Serum-free culture medium of NK (Natural Killer) cells
  • Serum-free culture medium of NK (Natural Killer) cells

Examples

Experimental program
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Effect test

Embodiment 1～4

[0029] The preparation process of the NK cell serum-free medium of Examples 1-4 is as follows:

[0030] Step 1: Obtain fatty acid-free albumin;

Embodiment 1

[0031] Embodiment 1 Get bovine serum albumin (the A1933 product purchased from SIGMA company) 1.5mmol, add water for injection and be made into 2000ml solution, add 50g gac, mix; Adopt the hydrochloric acid solution of 2mol / L to adjust the pH value to pH3.0; Place it in a water bath at 56°C for 30 minutes or place it overnight at 4°C; centrifuge (4000 rpm) to remove the activated carbon; then use 2mol / L NaOH solution to adjust the pH value to pH7.0; finally filter and sterilize to obtain the product of Example 1 fatty acid-free albumin solution;

Embodiment 2

[0032] Example 2 Take human serum albumin (purchased from Grifols (Grifols) company's human serum albumin (the product specification is 20% 50ml*10g / bottle, wherein the content of sodium octanoate is not more than 5mol / mol albumin)) 1.5 mmol, the specific operation process is the same as in Example 1, and the fatty acid-free albumin solution of Example 2 is obtained;

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Abstract

The invention relates to a serum-free culture medium of NK (Natural Killer) cells and a preparation method of the serum-free culture medium. The preparation method of the serum-free culture medium comprises the following steps: after loading fatty acid on fatty-acid-free albumin, adding an obtained albumin solution into a basic culture medium, so as to obtain the serum-free culture medium of the NK cells, which has more stable components and better function consistency and does not make a difference due to different species sources of the albumins or different processing batches. According tothe culture medium provided by the invention, polyinosinic-polycytidylic acid is also added; the polyinosinic-polycytidylic acid can be used for inducing the differentiation and maturing of immune cells or inducing the secretion of cell factors, such as IL-12 (Intedeukin-12), IL-15, TNFa (Tumor Necrosis Factor a) and IFN-r (Interferon-r), and can promote an activation and proliferation capabilityof the immune cells; the polyinosinic-polycytidylic acid also can be used for promoting an expression capability of membrane-bound IL-15Ra; the IL-15Ra can be tightly combined with the IL-15 and the IL-15 with the binding state has stronger activation promotion and proliferation promotion effects on the NK cells. The serum-free culture medium of the NK cells, provided by the invention, is more suitable for efficient culture of the NK cells and the number of times of proliferation is improved.

Description

technical field [0001] The invention relates to a NK cell serum-free medium and a preparation method thereof, belonging to the technical field of cell culture. Background technique [0002] Serum-free medium refers to a synthetic medium that can maintain the growth and reproduction of cells in vitro for a long time without adding serum, and can be used to cultivate immune cells, such as lymphocytes (for example, T lymphocytes, B cells, NK cells, etc.) , dendritic cells (also known as DC cells), monocytes / macrophages, etc. [0003] Generally speaking, the nutrients required for the survival and expansion of immune cells in vitro include: albumin, transferrin, insulin, amino acids, vitamins, sugars, lipids, inorganic salts, growth factors, etc. Among these components, albumin has the greatest impact on cell growth and survival, and is also the component with the most complex function and composition. In serum-free medium, the function of albumin is mainly determined by the f...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
CPCC12N5/0646C12N2500/24C12N2500/36C12N2500/40C12N2500/46C12N2500/90C12N2501/33C12N2501/998
Inventor 武宁谢志明
Owner 英普乐孚生物技术(上海)有限公司
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