METHOD FOR LONG-TERM CULTIVATION AND EXPANSION OF NK CELLS WITH HIGH VIABILITY AND FUNCTIONAL ACTIVITY Russian patent published in 2023 - IPC C12N5/783 

FIELD: biology; biotechnology; medicine.

SUBSTANCE: invention relates to a method for in vitro cultivation of natural killer (NK cells) in a nutrient medium with a combination of at least two cytokines: IL-2, IL-15, IL-18, IL-21 and K562-mb15-41BBL or K562-mb21-41BBL feeder cells. To implement this method, NK cells isolated from mononuclear cells (PBMC) of human peripheral blood are first added to the nutrient medium at a concentration of 1×10⁵ to 1×10⁶ per 1 ml of nutrient medium. After that, interleukins are added to the nutrient medium. Then feeder cells are added to the medium in ratios from 1:1 to 1:10. The entire process of culturing NK cells is 14–21 days, during which the replacement of the nutrient medium is carried out every 2–5 days from the beginning of cultivation. RPMI-1640 is used as a nutrient medium, in step (b) interleukins are added to the nutrient medium starting from day 0, in step (c) they start on day 5–8. Cultivation is carried out in a multi-gas incubator at 37°C in an atmosphere of 5% of CO₂ or 5% of CO₂ and 5-10% of O₂.

EFFECT: claimed method allows to obtain cell preparations with a high content of NK cells, namely up to 50×10⁷ cells, and allows to increase the functional activity of NK cells.

4 cl, 6 dwg, 5 ex