| CPC A61K 45/06 (2013.01) [A61K 40/15 (2025.01); A61K 40/428 (2025.01); C12N 5/0646 (2013.01); A61K 2239/48 (2023.05); C12N 2500/90 (2013.01); C12N 2501/2302 (2013.01)] | 7 Claims |
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1. A population of NK cells, wherein the population is obtained from the following steps:
collecting peripheral blood mononuclear cells from a healthy volunteer using Ficoll;
adding CD3 beads to the obtained peripheral blood mononuclear cells, and incubating the mononuclear cells and beads at 4° C. for 15 minutes;
adding a separation buffer to the mononuclear cells and the beads to obtain a suspension, and centrifuging the suspension at 300×g for 10 minutes;
removing a supernatant from the centrifuged suspension to obtain a residue, suspending the residue in a separation buffer, and adding the resulting suspension to LD Column wetted beforehand with the separation buffer;
collecting an eluate from the LD Column, and centrifuging the eluate at 500×g for 5 minutes;
removing a supernatant from the centrifuged eluate to obtain resultant cells, and suspending the resultant cells in NK medium I at a density of 5×105 cells/mL which are collected as primary NK cells, and
culturing the primary NK cells in a CO2 incubator at 37° C. under 5% CO2 atmosphere;
collecting the cells on day 14 of the culturing step as the population; and
wherein the NK medium I is COSMEDIUM 008 containing 5% of human AB type serum which is obtained by inactivation at 56° C. for 30 minutes.
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