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Preparation method of portable feeding layer for stimulating NK cell proliferation and differentiation

A technology of NK cells and feeder layer, which is applied in the field of immune cell experiments, can solve the problems of cancer cells, such as the risk, and achieve the effect of increasing the loss of manpower and facilitating cultivation

Inactive Publication Date: 2018-03-09
TIANJIN KANGTING BIOLOGICAL ENG GRP CO LTD
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Problems solved by technology

[0006] At present, the cell layer of NK cell culture is mainly based on K562 cell line transfection and improved feeder layer cells. Although this technology can stimulate the proliferation and maturation of NK cells in a large amount, even after radiation and other treatments, the nature of cancer cells still has risks.

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  • Preparation method of portable feeding layer for stimulating NK cell proliferation and differentiation
  • Preparation method of portable feeding layer for stimulating NK cell proliferation and differentiation

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Embodiment Construction

[0025] The present invention will be further described in detail below in conjunction with the accompanying drawings and through specific embodiments. The following embodiments are only descriptive, not restrictive, and cannot limit the protection scope of the present invention.

[0026] The present invention uses monocyte-induced macrophages as a substrate to construct a feeder layer of autologous cells. The method is simple and easy, and has obvious effects of stimulating proliferation and maturation of NK cells and other immune cells (cik cells, etc.) .

[0027] It is simple, easy to cultivate, and can provide raw materials for making DC vaccines while stimulating NK cells alone. In terms of safety, because no allogeneic heterologous cells are added, there will be no additional rejection caused by Symptoms such as fever.

[0028] A convenient feeder layer preparation method for stimulating NK cell proliferation and differentiation, which constitutes the core of the method ...

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Abstract

The invention relates to a preparation method of a portable feeding layer for stimulating NK cell proliferation and differentiation. The preparation method comprises the following steps: (1) collecting 50mL of peripheral blood; (2) separating mononuclear cells by adopting a ficoll density gradient centrifuging method; (3) separating lymphatic and mononuclear cells by utilizing an adherence methodand adhering for 1h; (4) adding factors into lymphocytes and culturing NK cells; (5) carrying out starving culture; after commonly culturing for 10h, collecting the cells. According to the preparationmethod of the portable feeding layer for stimulating the NK cell proliferation and differentiation, provided by the invention, in the aspect of safety, the feeding layer is a macrophage layer inducedby autologous and mononuclear cells and any allosome and heterogeneous cell is not added, so that symptoms including fever caused by extra rejection reaction and the like are not generated.

Description

technical field [0001] The invention belongs to the field of immune cell experiments, is mainly applicable to the differentiation and expansion of autologous NK cells, and can also be used for the preparation of other autologous immune cells, especially a convenient method for preparing a feeder layer for stimulating NK cell proliferation and differentiation. Background technique [0002] Natural killer cells (NK) are important immune cells in the body, not only related to anti-tumor, anti-virus infection and immune regulation, but also involved in the occurrence of hypersensitivity and autoimmune diseases in some cases. Since the killing activity of NK cells is not restricted by MHC and does not depend on antibodies, it is called natural killer activity. NK cells are rich in cytoplasm and contain larger azurophilic granules, and the content of granules is positively correlated with the killing activity of NK cells. The killing effect of NK cells occurs early after acting o...

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Application Information

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IPC IPC(8): C12N5/0783
CPCC12N5/0646C12N2501/22C12N2501/2304
Inventor 翟雷垒
Owner TIANJIN KANGTING BIOLOGICAL ENG GRP CO LTD
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