In-vitro culture method for improving activity of umbilical cord blood derived NK cells

Abstract

The invention discloses an in-vitro culture method for improving the activity of umbilical cord blood derived NK cells. The method adopts the following raw materials: a biosafety cabinet, a flow cytometer, a horizontal temperature control centrifuge, a sterilization pot, a CO2 incubator, an inverted microscope, a three-classification blood analyzer, a cell counter, a pipettor, refrigerators with the temperatures of -20 DEG C and 4 DEG C, a 75cm<2> culture bottle, a 225cm<2> culture bottle, a 2L culture bag, a 10ml transfer pipette, a 50ml efficient centrifuge tube, 15ml and 50ml centrifuge tubes, a 250ml centrifuge tube, medical sterile gloves, disposable sterile cap and sterile gloves, a serum-free culture medium LonzaX-VIVO15, recombinant interleukin 2, recombinant interleukin 15, a 0.4%trypan blue solution, physiological saline, human lymphocyte separation liquid, gamma interferon and recombinant interleukin 1[alpha]. According to the method, lentinan is added into related cytokines to culture the umbilical cord blood NK cells, so that the cost of culturing the NK cells by the method is not high, the proliferation speed is increased by 13-15 days, the proliferation multiple isincreased by about 270 times, the cell activity is high, and the cell phenotype is 91%.

Description

technical field [0001] The invention relates to the technical field of natural killer cells, in particular to an in vitro culture method for enhancing the activity of cord blood-derived NK cells. Background technique [0002] 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, and can recognize target cells and killing mediators. [0003] The exact source of NK cells is not very clear, it is generally believed to be directly derived from the bone marrow, and its development and maturation depends on the microenvironment of the bone marrow. In vitro experiments in mice and humans have shown that NK cells can also be induced by thymocytes cultured in the presence of cytokines such as IL-2 in vitro. Mouse spleen can promote the differentiation of NK cells under the induction of IL-3 in vi...

AI Technical Summary

Problems solved by technology

[0004] Due to the source and culture time of NK cells, the cost of culture is high and the speed of reproduction is slow, resulting in insufficient activity of the cultured cells. Therefore, we propose an in vitro culture method to improve the activity of cord blood-derived NK cells. solve the above problems

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