In-vitro amplification method of natural killer cells (NK)

Abstract

The invention discloses an in-vitro amplification method of natural killer cells (NK). The in-vitro amplification includes the steps of S1, culturing mononuclear cells separated from peripheral blood with a serum-free medium while adding in IFN-gamma to stimulate for a first preset time; S2, adding class-I cell factors in the serum-free medium for stimulation induction after the first preset time; S3, adding in class-II cell factors in the serum-free medium for induced amplification after culturing for a second preset time, and then performing cell fluid replacement and culturing for a third preset time; and S4, collecting the cells. According to embodiments of the invention, the natural killer cells cultured by the in-vitro amplification method have high killing action and high significant effect to target cells.

Description

technical field [0001] The invention relates to the technical field of cell culture, and more specifically, relates to an in vitro expansion method of natural killer cells. Background technique [0002] Natural killer cells (NK) are cytotoxic lymphocytes belonging to the lymphocyte lineage that contain perforin and granzyme granules. Its recognition of target cells is not restricted by MHC, it can directly kill tumor cells without prior sensitization, and can also secrete cytokines to regulate the functions of other immune cells. It is the main bearer of the body's natural immunity and the core of acquired cellular immunity. Regulatory cells play an important role in tumor immunity, anti-viral infection and removal of non-self cells. Studies have proved that natural killer cells are not only an important component of the natural immune system, but also have some characteristics of acquired immune cells. In the immune system, NK cells respond faster than T cells or B cells,...

AI Technical Summary

Problems solved by technology

[0003] However, due to the extremely low content of NK cells in human peripheral blood and the low distribution frequency (<10%) in tumor tissues, they only account for 5%-10% of mononuclear cells, which greatly limits the use of NK cells as adoptive immune cells. clinical application

For many years, people have been trying to achieve a large amount of NK cell expansion in vitro, but the current conventional technology is mainly to add a combination of factors such as IL-2, IL-12, IL-15 and γ-IFN to the in vitro culture system to promote the expansion of NK cells. NK amplification, after a period of culture, can only make NK cells amplify several times or ten times, and the purity is not ideal, so it cannot meet the practical application

Moreover, this method has a large demand for factors, high cost, and unstable culture effect, so it is not suitable for large-scale application in vitro.

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