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Culture method and serum-free culture medium composition of NK cells

A serum-free medium, NK cell technology, applied in the field of NK cell culture, to avoid uncertain effects, reduce costs, and increase final density

Active Publication Date: 2016-04-06
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The cell number, cell viability and killing activity of NK cells obtained by using existing media and culture methods still need to be further improved

Method used

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  • Culture method and serum-free culture medium composition of NK cells
  • Culture method and serum-free culture medium composition of NK cells
  • Culture method and serum-free culture medium composition of NK cells

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1, NK cell serum-free culture medium

[0035] In order to more clearly illustrate the components of the NK cell serum-free medium combination in this embodiment, they are listed below in the form of a table. Wherein Table 1 is the components and contents of the basal medium, and Table 2 is the induction components, proliferation components and activation components and their contents, and the induction components, proliferation components and activation components are added to the following contents respectively. In basal medium, induction medium, proliferation medium and activation medium can be obtained.

[0036] Table 1. Components and contents of basal medium

[0037]

[0038]

[0039] Induction component, proliferation component and activation component and content in table 2, embodiment 1

[0040]

Embodiment 2

[0041] Embodiment 2, NK cell serum-free culture medium

[0042] The components of the basal medium in this embodiment are shown in Table 3, and the induction components, proliferation components and activation components and their contents are shown in Table 4.

[0043] Table 3. Components and content of basal medium

[0044] components

Content (mg / L)

components

Content (mg / L)

L-Arginine

250.00

L-Aspartic Acid

20.00

L-Asparagine

40.00

Calcium Chloride Anhydrous

30.00

L-cystine dihydrochloride

55.15

potassium chloride

400.00

L-glutamic acid

20.00

Sodium chloride

6000.00

Glycine

10.00

glucose

2000.00

L-histidine

15.00

Reduced glutathione

1.00

L-Hydroxyproline

20.00

Vitamin B12

0.005

L-isoleucine

50.00

L-Glutamine

300.00

L-leucine

50.00

biotin

0.30

L-Lysine Hydrochloride

40.00

...

Embodiment 3

[0048] Embodiment 3, NK cell serum-free culture medium

[0049] The components of the basal medium in this embodiment are shown in Table 5, and the induction components, proliferation components and activation components and their contents are shown in Table 6.

[0050] Base culture medium component and content in table 5, embodiment 3

[0051]

[0052]

[0053] Induction component, proliferation component and activation component and content in table 6, embodiment 3

[0054]

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Abstract

The invention discloses a culture method and serum-free culture medium composition of NK cells. The serum-free culture medium composition of the NK cells comprises an induction culture medium, a proliferation culture medium and an activation culture medium; the induction culture medium comprises a basal culture medium and an induction component, the proliferation culture medium comprises a basal culture medium and a proliferation culture component, and the activation culture medium comprises a basal culture medium and an activation component. The culture method of the NK cells comprises the step of performing cell culture by sequentially using the induction culture medium, the proliferation culture medium and the activation culture medium in stages. The serum-free culture medium composition of the NK cells comprises the induction culture medium, the proliferation culture medium and the activation culture medium which are designed for the different influence requirements of the induction, proliferation and activation periods of the NK cells, and the cultured NK cells are superior to those cultured through an existing technology on the aspects such as the total cell number, the proliferation speed, the amplification times and the killing activity on tumor cells.

Description

technical field [0001] The invention relates to the field of NK cell culture, in particular to a NK cell culture method and serum-free medium combination. Background technique [0002] Natural killer cells (NK cells) are the main cells of the body's innate immunity. They are non-specific in recognizing target cells, and participate in the recognition process of NK cells through the action of intercellular adhesion molecule-1 (ICAM-1) on the surface of target cells. In addition, NK cells kill target cells by secreting perforin, NK cytotoxic factors and TNF. Since the killing activity of NK cells is not restricted by MHC and does not depend on antibodies, it is called natural killer activity. Therefore, NK cells play an important role in cellular immunotherapy. [0003] The NK cells in the existing cellular immunotherapy are mainly separated from the lymphocytes in the blood, cultured in vitro, stimulated by various cytokines, induced and expanded, and then a certain amount ...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
Inventor 王一飞陈海佳葛啸虎应杰王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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