Efficient in-vitro amplification method of human natural killer cells

Abstract

The invention discloses an efficient in-vitro amplification method of human natural killer (NK) cells. The efficient in-vitro amplification method includes the steps that CD16 monoclonal antibodies, ATG and IL-2 are added into a processed culturing system, incubation is carried out in an incubator at the temperature of 37 DEG C, counting and solution changing are carried out, and fresh IL-2 is added; a culture solution is changed at regular intervals, culturing continues for a proper time, and the human NK cells are obtained. The efficient in-vitro amplification method has the advantages that the NK cells can be efficiently amplified through the ATG and the CD16 antibodies in a short time without being subjected to separation of magnetic beads and irritation of tumor modification cells, production cost is greatly saved, and operation is easy and convenient; the number of amplification times of the NK cells is one thousand or more, and the final proportion ranges from 80% to 93%; production efficiency is high, and the efficient in-vitro amplification method can be entirely applied to large-scale clinical researching and the like; the growth process of the NK cells is completed under the GMP condition, sterile operation is guaranteed, various kinds of security detection are carried out at the final production stage, and the efficient in-vitro amplification method can be applied to clinical researching.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a method for high-efficiency expansion of human natural killer cells in vitro. Background technique [0002] Since the biological activity of NK cells was discovered in the 1880s, the research on NK cells and its application have been continuously carried out. Clinical trials at home and abroad have proved that NK cells are effective in treating hematological tumors, and it is expected to become one of the ways to completely cure hematological tumors. However, the clinical application of NK cells is also limited by their limited number. NK cells only account for 5%-15% of human peripheral blood lymphocytes. Therefore, the high-efficiency expansion of NK cells in vivo and in vitro is a matter of great concern to researchers and one of the bottlenecks in the widespread clinical application of NK cells. Only by solving the limitation of the number of NK cells, can the clinical...

AI Technical Summary

Problems solved by technology

[0005] The above methods of scholars at home and abroad have allowed NK cells to be expanded in large quantities, but the use of magnetic beads for separation has greatly increased the research cost, and with the help of K562, Wilms tumor cell line HFWT cells and other genetically modified tumor cells to stimulate NK expansion, Its security is at risk

Although the research methods of other scholars are easy to operate, the amplification efficiency is low.

At present, there are commercial NK cell isolation kits with an isolation purity of more than 95%, but they are expensive and difficult to meet the requirements of large-scale experiments and clinical tumor immunotherapy. Therefore, NK cell culture with simple operation and high expansion efficiency has been studied. technology is necessary

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