Culture method and application of NK cells

Abstract

The invention discloses a culture method and application of NK cells, which belong to the technical field of cell culture. The culture method of the NK cells comprises the following steps of (1) antibody coating, (2) separating umbilical cord blood mononuclear cells and preparing serum, (3) activating and culturing the NK cells, and (4) multiplication culture of the NK cells. According to the separation culture method provided by the invention, the problem that lymphocytes are difficult to obtain is solved, erythrocyte pollution is eliminated in the initial stage of culture, the purity of NK cells harvested in the later stage can reach 90% or above, the defect that the culture purity of NK cells of tumor patients is low is overcome, and the culture method has huge advantages in the aspect of application of allogeneic NK cells.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a method for culturing NK cells and an application thereof. Background technique [0002] NK cells, also known as natural killer cells (natural killer cells, NK cells), are important immune cells in the body, responsible for killing abnormal cells such as aging, virus infection, and tumors. This natural killing activity does not require antigen sensitization and has no MHC restrictive. It can recognize and attack foreign viruses, cancer cells and abnormal cells by itself. Once activated, it will directly attack target cells by releasing lethal media such as granzyme and perforin and secreting a large number of cytokines, or through antibody-dependent cytotoxicity (ADCC) effect) to kill tumor cells, or induce apoptosis of target cells by inducing Fas / Fasl and tumor necrosis factor-related apoptosis-inducing ligand (Trail) molecules. Adoptive immunotherapy is to ...

AI Technical Summary

Problems solved by technology

However, the expansion methods at home and abroad are different and cumbersome. Therefore, the acquisition of high-quality NK cells is still the key to restricting their clinical application. It is a huge task to generate a large number of NK cells that meet GMP (Good Manufacturing Practice) standards for the treatment of patients. challenge

[0004] CN108251376A discloses an artificial antigen-presenting cell and its construction method and its application in the expansion of chimeric antigen receptor T cells. However, research has found that the constructed artificial antigen-presenting cells cannot expand NK cells, and The artificial antigen-presenting cells are relatively complex and costly

[0005] CN108300693A discloses a method for amplifying natural killer cells in vitro. The NK cells amplified by the method have poor antitumor activity. Only when NK cells:tumor target cells=10:1 can a good antitumor effect be produced. Good for clinical application

At the same time, the study found that the expanded NK cells had poor tumoricidal activity under hypoxic conditions.