A novel cytokine combination for CIK cell culture

A factor combination and cytokine technology, applied in the field of cells, can solve the problems of low killing activity and poor CIK cell proliferation effect, and achieve the effects of strong killing activity, high cell surface antigen content, and large amplification multiple.

Inactive Publication Date: 2019-05-03
深圳光彩生命工程技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a new combination of factors for cultivating CIK cells. The technical problem to be solved is: the proliferation effect of CIK cells is poor and the killing activity is low, so it is necessary to solve these problems

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] A new factor combination for culturing CIK cells, which consists of the following raw materials: IFN-λ 600U / mL, 70U / mL IL-1α, 12ng / ml OKT-3, 1000U / ml IL-2 cytokines, 200U / mL interference Glycerin-γ, 10ng / mL CD3 stimulating monoclonal antibody, 100U / mL interleukin-2, 0.05mg / mL lentinan and serum-free basal medium.

[0018] In the above embodiments, serum is also included, and the content of the serum is 50 U / mL.

[0019] In the above examples, the serum-free basal medium is RPMI-1640 medium provided by RPMI (Roswell Park Memorial Institute).

[0020] Lentinan is added to the CIK cell culture factor combination provided by the present invention, and lentinan can promote PBMCs to produce lymphocyte activating factors and release helper T cell factors, thereby promoting the proliferation of CIK cells, and then making the cultured CIK cells exhibit Large amplification factor, strong killing activity and high cell surface antigen content.

[0021] At the same time, the pres...

Embodiment 2

[0023] A new factor combination for culturing CIK cells, which is composed of the following raw materials: IFN-λ 1300U / mL, 130U / mL IL-1α, 48ng / ml OKT-3, 100U / ml IL-2 cytokine, 1000U / mL interference Glycerin-γ, 80ng / mL CD3 stimulating monoclonal antibody, 600U / mL interleukin-2, 0.3mg / mL lentinan and serum-free basal medium.

[0024] In the above embodiments, serum is also included, and the content of the serum is 2000 U / mL.

[0025] In the above examples, the serum-free basal medium is RPMI-1640 medium provided by RPMI (Roswell Park Memorial Institute).

[0026] Lentinan is added to the CIK cell culture factor combination provided by the present invention, and lentinan can promote PBMCs to produce lymphocyte activating factors and release helper T cell factors, thereby promoting the proliferation of CIK cells, and then making the cultured CIK cells exhibit Large amplification factor, strong killing activity and high cell surface antigen content.

[0027] At the same time, the...

Embodiment 3

[0029] A new factor combination for culturing CIK cells, which consists of the following raw materials: IFN-λ 900U / mL, 100U / mL IL-1α, 30ng / ml OKT-3, 550U / ml IL-2 cytokines, 600U / mL interference Glycerin-γ, 45ng / mL CD3 stimulating monoclonal antibody, 260U / mL interleukin-2, 0.15mg / mL lentinan and serum-free basal medium.

[0030] In the above embodiments, serum is also included, and the content of the serum is 1200 U / mL.

[0031] In the above examples, the serum-free basal medium is RPMI-1640 medium provided by RPMI (Roswell Park Memorial Institute).

[0032] Lentinan is added to the CIK cell culture factor combination provided by the present invention, and lentinan can promote PBMC to produce lymphocyte activating factors and release helper T cell factors, thereby promoting the proliferation of CIK cells, and then making the cultured CIK cells exhibit Large amplification factor, strong killing activity and high cell surface antigen content.

[0033] At the same time, the pre...

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Abstract

The invention relates to a novel cytokine combination for CIK cell culture. The cytokine combination includes 600-1300 U / mL of IFN-gamma, 70-130 U / mL of IL-1alpha, 12-48 ng / mL of OKT-3, 100-1000 U / mLof IL-2 cytokine, 200-1000 U / mL of interferon gamma, 10-80 ng / mL of a CD3 exciting type monoclonal antibody, 100-600 U / mL of interleukin-2, 0.05-0.3 mg / mL of lentinan and a serum-free base medium. Compared with the prior art, CIK cells cultured by utilizing the cytokine combination show a high amplification time, strong killing activity and high cell surface antigen contents because the cytokine combination contains the lentinan and can promote CIK cell proliferation. Through optimizing the culture cytokine combination used in a CIK cell culture process, CIK cell proliferation is further promoted and tumor killing activity of the CIK cells is enhanced.

Description

technical field [0001] The invention relates to the field of cell technology, in particular to a new combination of factors for cultivating CIK cells. Background technique [0002] CIK cells (cytokine induced killer, cytokine-induced killer cells) are a new type of immunocompetent cells, which are autologous peripheral blood mononuclear cells, a type of CD3+ and CD56+ membrane protein molecules produced by co-induction and culture of various cytokines Co-expressed killer cells. CIK cells have anti-tumor activity and non-MHC tumor-killing properties. They are effector cells with the characteristics of fast proliferation, high tumor-killing activity, and broad tumor-killing spectrum. [0003] At present, the common method for culturing CIK cells in vitro is to culture peripheral blood mononuclear cells (PBMC) in serum-free medium supplemented with corresponding cytokines, and induce PBMCs to become CIK cells under the stimulation of cytokines. But when adopting this method t...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C12N5/0783
Inventor陈阳甘露
Owner深圳光彩生命工程技术有限公司