Culture method and serum-free culture medium composition of NK cells
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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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The cell number, cell viability and killing activity of NK cells obtained b
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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 ...
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