	US 12,291,721 B2
	Activated lymphocytes comprising cytokine-induced killer cells and preparation method therefor
Gyou Chul Jung, Yongin-si (KR); Dong Young Kim, Yongin-si (KR); and Ji Min Lee, Yongin-si (KR)
Assigned to GREENCROSSCELL, Seoul (KR)
Appl. No. 17/434,814
Filed by GREENCROSSCELL, Seoul (KR)
PCT Filed May 4, 2020, PCT No. PCT/KR2020/005896 § 371(c)(1), (2) Date Aug. 30, 2021, PCT Pub. No. WO2020/231058, PCT Pub. Date Nov. 19, 2020.
Claims priority of application No. 10-2019-0057531 (KR), filed on May 16, 2019; and application No. 10-2019-0168127 (KR), filed on Dec. 16, 2019.
Prior Publication US 2022/0160763 A1, May 26, 2022
Int. Cl. C12N 5/078 (2010.01); A61K 40/10 (2025.01); A61K 40/42 (2025.01); A61P 35/00 (2006.01); C12N 5/0783 (2010.01)

CPC C12N 5/0634 (2013.01) [A61K 40/10 (2025.01); A61K 40/42 (2025.01); A61P 35/00 (2018.01); C12N 5/0646 (2013.01); A61K 2239/31 (2023.05); A61K 2239/38 (2023.05); A61K 2239/46 (2023.05); C12N 2501/2302 (2013.01); C12N 2501/515 (2013.01)]

1 Claim

1. A method for preparation of activated lymphocytes comprising CD8⁺CD56⁺NKG2D⁺ cells, the method comprising culturing lymphocytes isolated from peripheral blood in a medium in three steps,

wherein a medium in a first step contains an anti-CD3 antibody, interleukin-2 and fetal bovine serum (FBS), a medium in a second step and a third step contains interleukin-2 and FBS and does not contain the anti-CD3 antibody, and

wherein a concentration of the anti-CD3 antibody is from 1 to 10 μg/ml, a concentration of interleukin-2 in each of the first, the second and the third step is from 100 to 800 U/ml, and a concentration of FBS in the first step is from 5 to 15% by volume, a concentration of FBS in the second step is from 0.1 to 1% by volume, and a concentration of FBS in the third step is from 0.1 to 2% by volume.