Culture medium and method used for in-vitro large-scale amplification of NK cells

Abstract

The invention relates to a culture medium and a method used for in-vitro large-scale amplification of NK cells. By volume percent, the culture medium comprises 100% of a lymphocyte serum-completely-free culture medium or a mixture including 1-95 % of DMEM / F12 culture medium and 5-99 % of the lymphocyte serum-completely-free medium, and human albumin and interleukin 2, wherein the concentration ofthe albumin is 0.1-2 g / L, and the concentration of the interleukin 2 is 100-600 [mu] g / mL. With the culture medium and the preparation method provided by the invention, the rapid large-scale amplification within a short period of the NK cells can be realized, the cost is low, and the amplification has very high lethality.

Description

technical field [0001] The invention relates to the technical field of immunology, in particular to a culture medium and method for large-scale expansion of NK cells in vitro. Background technique [0002] Natural killer cells are important immune cells in the body and play an important role in anti-tumor and anti-viral infection immunity. Since NK cell killing activity has no MHC limit, it is called natural killer cell. The target cells of NK cells are mainly tumor cells, virus-infected cells, some self-organized cells (such as blood cells) and parasites, etc. Therefore, NK cells are an important part of the body's anti-tumor and anti-infection immunity. Due to these characteristics, NK cells have broad application prospects in cellular immunotherapy. NK cells activated in vitro are highly toxic and can be used as therapeutic preparations for immune cells. Human in vitro and animal experiments have confirmed the possibility of its treatment of various cancers: the use of...

AI Technical Summary

Problems solved by technology

Due to the low content of NK cells in human peripheral blood (accounting for about 5% to 10% of lymphocytes), the cost of isolating pure NK cells from peripheral blood is expensive, and the isolated cells are in an inactive state, which limits the its clinical application

To achieve the therapeutic effect, the required effective NK cell number in a single batch should be at least 5×109 (and the purity should reach more than 80%). At present, it is reported that the serum-free culture system is used to pass IL-2, IL-21 and Stimulation and culture of related combination factors cannot achieve high activity, high purity and high expansion multiple of NK cells, and its expansion ability can reach up to 300 times, and there are safety risks in clinical application

Scale expansion is achieved through the autologous serum culture system, and there is no report of obtaining a single culture system, the number of cells expanded in a single batch is greater than 5×109 NK cells, and the purity is greater than 90%

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