Method for efficiently amplifying NK cells in vitro by factor method

Abstract

The invention discloses the technical field of NK cell amplification. The invention relates to a method for efficiently amplifying NK cells in vitro by a factor method. The method comprises the following steps: carrying out cell separation, cell inoculation and activation, liquid exchange and cell amplification; according to the invention, a CD16 monoclonal antibody and a NKP46 monoclonal antibodyare matched for use, the OK432 is used, and the IL-15 and the hydrocortisone are matched for use; the method is characterized by directly culturing PBMCs by utilizing cured Anti-CD16mAb and NKP46 monoclonal antibodies; the culture bottle does not need to be pre-coated; OK432 is used as a biological adjuvant; NK cells are amplified in vitro under the combined action of hydrocortisone, IL-2 and IL-15; and hydrocortisone is an immunosuppressive agent. When used with IL-15 at the same time; hydrocortisone can be used for promoting proliferation of IL-15 activated NK cells; according to the present invention, the efficient NK cell in vitro amplification method is created, the CD3-CD16+ / CD56+ cell expression rate of the NK cell prepared through the method is up to 60% or more, and after 15-18 days of culture, the NK cell can be amplified by 1000 or more.

Description

technical field [0001] The invention relates to the technical field of NK cell expansion, and the specific field is a method for efficiently expanding NK cells in vitro by factor method. Background technique [0002] NK cells, that is, natural killer cells, are important immune cells in the body. They are not only related to anti-tumor, anti-viral infection and immune regulation, but also participate in the occurrence of hypersensitivity and autoimmune diseases in some cases. They can recognize target cells, Lethal medium. Natural killer cells are derived from bone marrow lymphoid stem cells. Their differentiation and development depend on the microenvironment of bone marrow and thymus, and are mainly distributed in bone marrow, peripheral blood, liver, spleen, lung and lymph nodes. NK cells are different from T and B cells. They are a type of lymphocytes that can non-specifically kill tumor cells and virus-infected cells without prior sensitization. Since the killing activ...

AI Technical Summary

Problems solved by technology

[0004] 1. More protein active factors (such as IL-4, IL-2, IL-15, IL-18, IL-21, etc.) need to be used. These factors are relatively expensive and difficult to store and pack;

[0005] 2. It is necessary to cover the culture bottle one day in advance, which increases the uncertainty of the culture. If the culture plan is canceled temporarily, the pre-prepared materials will not be available, resulting in waste;

[0006] 3. Need to use magnetic beads to sort and purify the steps, increasing the difficulty and cost of operation;

[0007] 4. The existing technology often uses inactivated feeder cells for joint culture, but the feeder cells themselves are transformed leukemia cells, and there are certain safety risks in use

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