A kind of cik cell culture medium

A culture medium and basal medium technology, applied in the direction of cell culture active agent, culture process, animal cells, etc., can solve the problems of poor CIK cell activity and proliferation ability, and the number of CIK cells is not ideal, so as to improve the activity and proliferation ability , Avoid reducing the concentration too fast, and enhance the effect of cross-linking

Inactive Publication Date: 2019-12-20
DONGGUAN BOALAI BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the CIK cells cultured in the existing medium have the problem of poor activity and proliferation ability, and the finally obtained CD3 + CD56 + The number of CIK cells is not ideal

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The present embodiment provides a kind of CIK cell culture medium, and described CIK cell culture medium comprises DMEM medium and the additive that is added in DMEM medium, by final concentration, described additive comprises the component of following content: polylysine acid 25μg / mL, BCG polysaccharide nucleic acid 10mg / mL, sodium selenite 0.035μmol / mL, 4-hydroxyethylpiperazineethanesulfonic acid 6μg / mL, reduced glutathione 3μg / L, adenosine Ammoniase 9µg / mL, IFN-γ sustained-release microspheres 2.5mg / mL, IL-2 sustained-release microspheres 1.75mg / mL, autologous plasma 5mL / L.

[0033] Preferably, the preparation method of the IL-2 sustained-release microspheres comprises the following steps:

[0034] A, adding IL-2 and polyethylene glycol to the aqueous solution that has been added with mannitol and zinc sulfate in advance, after stirring evenly, a mixture A is obtained, the concentration of IL-2 in the mixture A is 2wt%, and the concentration of polyethylene glycol ...

Embodiment 2

[0055] The present embodiment provides a kind of CIK cell culture medium, and described CIK cell culture medium comprises basal medium and the additive that is added in the basal medium, and in terms of final concentration, described additive comprises the component of following content: Polylysine acid 20μg / mL, BCG polysaccharide nucleic acid 15mg / mL, sodium selenite 0.03μmol / mL, 4-hydroxyethylpiperazineethanesulfonic acid 8μg / mL, reduced glutathione 2μg / L, adenosine Ammoniase 12µg / mL, IFN-γ sustained-release microspheres 2mg / mL, IL-2 sustained-release microspheres 2mg / mL, autologous plasma 4mL / L.

[0056] Preferably, the preparation method of the IL-2 sustained-release microspheres comprises the following steps:

[0057] A. Add IL-2 and polyethylene glycol to the aqueous solution pre-added with mannitol and zinc sulfate, and stir evenly to obtain mixture A. The concentration of IL-2 in the mixture A is 1.2wt%, polyethylene glycol The concentration is 12 wt%;

[0058] B. Ad...

Embodiment 3

[0079] The present embodiment provides a kind of CIK cell culture medium, described CIK cell culture medium comprises DMEM medium and the additive that is added in the basal medium, by final concentration, the described additive comprises the component of following content: polylysine acid 30μg / mL, BCG polysaccharide nucleic acid 5mg / mL, sodium selenite 0.04μmol / mL, 4-hydroxyethylpiperazineethanesulfonic acid 5μg / mL, reduced glutathione 4μg / L, adenosine Ammoniase 8µg / mL, IFN-γ sustained-release microspheres 3mg / mL, IL-2 sustained-release microspheres 1.5mg / mL, autologous plasma 6mL / L.

[0080] Preferably, the preparation method of the IL-2 sustained-release microspheres comprises the following steps:

[0081] A. Add IL-2 and polyethylene glycol to the aqueous solution pre-added with mannitol and zinc sulfate, and stir evenly to obtain mixture A. The concentration of IL-2 in the mixture A is 2.5wt%, polyethylene glycol The concentration is 7wt%;

[0082] B. Add polyglycolic a...

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Abstract

The invention relates to the biotechnology field of biotechnolog, in particular to a CIK (cytokine induced killer cell) medium, which comprises a basic medium and an additive added into the basic medium. In terms of a final concentration, the additive comprises the following components: 20-30microg / mL polylysine, 5-15mg / mL BCG-polysaccharide, 0.03-0.04micromol / mL sodium selenite, 5-8microg / mL 4-hydroxyethylpiperazine ethane sulfonic acid, 2-4microg / L reduced glutathione, 8-12microg / mL adenosine deaminase, 2-3mg / mL IFN-gamma sustained release microsphere, 1.5-2mg / mL IL-2 sustained release microspheres, and 4-6mL / L autologous plasma. Through intercoordination and combined action of all the components, the CIK medium provided by the invention can provide sufficient nutrition and good environment needed by cell growth and proliferation, and can improve the activity and multiplication capacity of immune cells.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a CIK cell culture medium. Background technique [0002] Cell adoptive immunotherapy is a biological treatment method in which immune cells induced in vitro with strong tumoricidal activity are returned to patients. It not only has the ability to kill residual cancer cells in the body, but also enhances the immunity of the host. One of the important means of biological therapy. Cytokine induced killer cells (CIK) are a group of heterogeneous cells obtained by co-culturing and stimulating human peripheral blood mononuclear cells (PBMCs) with various cytokines. CD3 + CD56 + Double-positive cells have both the strong anti-tumor activity of T lymphocytes and the characteristics of non-major histocompatibility complex (MHC)-restricted tumor killing, so CIK cells are considered to be the first choice for adoptive immunotherapy against tumor cells . [0003] CIK cells can proliferate ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0638C12N2500/12C12N2500/30C12N2500/50C12N2501/2302C12N2501/24C12N2501/73C12N2501/90C12N2501/998
Inventor 罗天恩
Owner DONGGUAN BOALAI BIOLOGICAL TECH CO LTD
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