Application of clinical applicable culture system for efficient amplification of NK cells

Abstract

The invention relates to application of a clinical applicable culture system for efficient amplification of NK cells. The invention specifically provides application of a serum-free culture system for amplification of NK cells. The culture system comprises the following components: 1640 medium, sodium bicarbonate, human serum albumin, transferrin, linoleic acid, oleic acid, palmitic acid, sodium pyruvate, human recombinant insulin, non-essential amino acids, 2-mercaptoethanol and interleukin-2. The invention also provides application of a culture system containing 1-12 vol.% of human autoserum and for amplification of NK cells. The culture system provided by the invention has simple composition, avoids the risks of animal ingredients in animal serum on cell treatment and uncertainty of cell culture caused by the uncertain components in the animal serum, and increases yield of NK cell. The obtained activated NK cells have significant tumor killing effect. Therefore, the system can not only be applied in scientific research but also be used in clinical treatment.

Description

technical field [0001] The invention relates to the field of biology, in particular, it relates to the application of a culture system for high-efficiency expansion of NK cells suitable for clinical use. Background technique [0002] NK cells (natural killer cells) were discovered in 1975 and belong to the lymphocyte lineage. They are cytotoxic lymphocytes containing perforin and granzyme granules, and are important immune cells in the body. NK cells are derived from bone marrow and belong to large granular lymphocytes in morphology, accounting for 5%-10% of the total number of peripheral blood lymphocytes. Their recognition of target cells is not restricted by MHC, and they can directly kill tumor cells without prior sensitization. It 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 regulatory cell of acquired cellular immunity. It plays an important role in tumor immunity, anti-vir...

AI Technical Summary

Problems solved by technology

[0005]① NK cells only account for 5%-20% of normal peripheral blood lymphocytes, and the content is very small. The cost of isolating NK cells from peripheral blood is high, and the isolated NK cells are generally in an inactive state, which largely limits their clinical application

Moreover, K562 cells are erythroleukemia cells. This method is complicated to operate and the cell yield is low, which reduces the safety of clinical use to a certain extent.

[0007]③ The composition of animal serum is complex and contains components harmful to cells, such as polyamine oxidase, which can react with polyamines from highly proliferative cells to form cytotoxic Polyspermine, complement, antibodies, bacterial toxins, etc., will affect cell growth and even cause cell death; different animals have different serum origins and batches, and their components cannot be consistent; Lead to experimental failure or unreliable experimental results; in large-scale production, the source of serum is difficult and expensive

Not suitable for clinical cell therapy system

[0008]④ Some NK cell culture medium products need to use a large amount of antibody-coated culture consumables during use, and the antibody is expensive, which increases the cost

However, the composition of the above medium is very complex

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