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Culture medium efficiently amplifying autologous NK cells and cultural method

A technology for NK cells and cell culture, applied in the field of cell biology, can solve the problems of NK cell culture technology and medium production technology that need to be broken through, and achieve the effect of being conducive to large-scale production, significant curative effect, and significant anti-tumor effect.

Active Publication Date: 2014-06-11
康思葆(北京)生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main reason is that the NK cell culture technology and culture medium production technology need to be broken through

Method used

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  • Culture medium efficiently amplifying autologous NK cells and cultural method
  • Culture medium efficiently amplifying autologous NK cells and cultural method
  • Culture medium efficiently amplifying autologous NK cells and cultural method

Examples

Experimental program
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Embodiment 1

[0026] A medium for efficiently expanding autologous NK cells, comprising a cell culture medium and an additive factor added to the cell culture medium for efficiently expanding autologous NK cells, the additive factor comprising interleukin 2 (IL-2), interleukin 15 (IL-15), interleukin 7 (IL-7), interleukin 12 (IL-12), tumor necrosis factor alpha (TNFα) and anti-CD3 antibody.

[0027] The medium can be common cell culture medium such as RPMI-1640 medium, X-VIVO serum-free medium or GT-T551 medium.

[0028] Further, the medium for cell culture contains 175-1750 IU / ml interleukin-2.

[0029] Further, the medium for cell culture contains 10-100 ng / ml interleukin-15.

[0030] Further, the medium for cell culture contains 10-100 ng / ml interleukin-7.

[0031] Further, the medium for cell culture contains 10-100 ng / ml interleukin-12.

[0032] Further, the medium for cell culture contains 10-100 ng / ml tumor necrosis factor α.

[0033] Further, the medium for cell culture and the ...

Embodiment 2

[0038] This example is an optimization scheme based on Example 1, and provides a medium and a culture method for efficiently expanding autologous NK cells. For the parts in this embodiment that are the same as those in Embodiment 1, please refer to the content disclosed in Embodiment 1 for understanding, and the content disclosed in Embodiment 1 should also be regarded as the content of this embodiment, and will not be described repeatedly here.

[0039] The culture method of the expansion autologous NK cell of the present embodiment is:

[0040] 1) Cell stimulating factor coating

[0041] Dilute CD3mAb (10 ug / ml) with sterile PBS, take 5 mL and add to 75 cm 2 In the cell culture bottle, lightly tap to make the liquid cover the bottom of the bottle, then lay the cell culture bottle flat, leave it at room temperature overnight, and then transfer it to a 4 ℃ refrigerator for storage until use.

[0042] 2) Preparation of peripheral blood mononuclear cells (PBMCs)

[0043]Colle...

Embodiment 3

[0056] This example is an optimization scheme based on Example 1, which provides a medium for efficiently expanding autologous NK cells. For the parts in this embodiment that are the same as those in Embodiment 1, please refer to the content disclosed in Embodiment 1 for understanding, and the content disclosed in Embodiment 1 should also be regarded as the content of this embodiment, and will not be described repeatedly here. Please refer to Example 2 for the cultivation method.

[0057] The medium used for cell culture in this embodiment is RPMI-1640 medium, and the composition of the medium is:

[0058] RPMI-1640 medium 10 ml

[0059] Interleukin 2 (IL-2) 20000 IU

[0060] Interleukin-15 (IL-15) 500 ng

[0061] Interleukin 7 (IL-7) 500 ng

[0062] Interleukin 12 (IL-12) 500 ng

[0063] Tumor necrosis factor alpha (TNFα) 500 ng.

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Abstract

The invention relates to a culture medium efficiently amplifying autologous NK cells and a culture method. The culture medium internally contains interleukin 2(IL-2), interleukin 15(IL-15), interleukin 7(IL-7), interleukin 12(IL-12), tumor necrosis factor alpha (TNFalpha) and CD3-antibody, can efficiently amplify and activate NK cells so as to obtain a large quantity of highly active immune cells with majority of NK cells to be transferred back to human bodies, and has remarkable curative effect in anti-tumor, anti-virus infection and immune adjustment related diseases. The culture medium efficiently amplifying autologous NK cells comprises the culture medium for cultivating cells and added factors added into the culture medium for cultivating cells, is used for cultivating and amplifying a large quantity of autologous activated NK cells, and is characterized in that the added factor comprises interleukin 2(IL-2), interleukin 15(IL-15), interleukin 7(IL-7), interleukin 12(IL-12), tumor necrosis factor alpha (TNF alpha) and CD3-antibody.

Description

technical field [0001] The invention belongs to the field of cell biology, in particular to a culture medium and a culture method for efficiently expanding autologous NK cells. Background technique [0002] In my country, malignant tumors have become the first cause of death among residents. Tumor biological immunotherapy is a treatment method other than surgery, radiotherapy and chemotherapy, and has become one of the most promising measures for human beings to fight tumors. NK cell adoptive immunotherapy is a kind of tumor biological immunotherapy. Its principle is to apply modern biomedical technology to activate and expand human NK cells in vitro, and then reinfuse them to adjust and stimulate the body's anti-tumor immune system. thereby killing tumor cells. At the same time, NK cell adoptive immunotherapy also has a certain effect on viral or bacterial infections, immune regulation-related diseases, and anti-aging. [0003] NK cells are considered to be the body's ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
Inventor 吴海涛周丽丽
Owner 康思葆(北京)生物技术有限公司
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