METHOD FOR EXPANSION OF HUMAN NK CELLS USING FEEDER CELLS Russian patent published in 2022 - IPC C12N5/783 

FIELD: medicine.

SUBSTANCE: invention relates to cell biology and medicine, in particular to a method for expansion of NK cells derived from human peripheral blood. To implement the method, mononuclear cells (MNCs) are first isolated from human peripheral blood by gradient centrifugation. MNCs are brought to a concentration of 5 million/ml, seeded into a Petri dish or culture flask in a pure RPMI medium and incubated for 60 minutes in a CO₂ incubator at 37°C and 5% CO₂. After that, the loose NK cells are collected, centrifuged at 1000 rpm for 5 minutes and brought to the desired concentration, then in a CO₂ incubator in a nutrient medium RPMI-1640 with IL-2, 300 U/ml and 10% FBS. Then, NK cell proliferation is stimulated using feeder cells previously obtained by irradiation of the K562 cell line with a dose of 100 Gy in a full growth medium RPMI-1640 containing IL-2 in a CO₂ incubator at 37°C and 5% CO₂. Moreover, feeder cells are added to NK cells three times: on day 0 of cultivation in a ratio of 1:1, on day 7, 2:1 and on day 14, 5-10:1. Every 48 hours, the nutrient medium is updated with the addition of IL-2, 300 U/ml. Feeder cells are used immediately after receiving or storing in liquid nitrogen with a cryoprotector consisting of 10% dimethyl sulfoxide (DMSO) and 90% embryonic calf serum (FBS). The duration of the expansion is 21 days.

EFFECT: present invention makes it possible to increase the proliferative potential of NK cells in vitro.

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