US 12,435,310 B2
Expansion culture method for human-derived natural killer cells by using HDAC inhibitor
Kyung-Mi Lee, Seoul (KR); Seon Ah Lim, Seoul (KR); and Cassian Yee, Seoul (KR)
Assigned to Korea University Research and Business Foundation, Seoul (KR)
Appl. No. 17/055,678
Filed by KOREA UNIVERSITY RESEARCH AND BUSINESS FOUNDATION, Seoul (KR)
PCT Filed May 13, 2019, PCT No. PCT/KR2019/005721
§ 371(c)(1), (2) Date Nov. 16, 2020,
PCT Pub. No. WO2019/221463, PCT Pub. Date Nov. 21, 2019.
Claims priority of application No. 10-2018-0055858 (KR), filed on May 16, 2018.
Prior Publication US 2021/0198627 A1, Jul. 1, 2021
Int. Cl. C12N 5/0783 (2010.01)
CPC C12N 5/0646 (2013.01) [C12N 2501/2302 (2013.01); C12N 2501/999 (2013.01)] 1 Claim
 
1. A culture method for expansion of natural killer cells, the method consisting of:
(a) co-culturing natural killer cells with Jurkat cell line from Korea Cell Line Bank and EBV-LCL cell line from Korea Cell Line Bank, where each of the Jurkat cell line and the EBV-LCL cell line was irradiated with 100 Gy and at the natural killer cell lines: the Jurkat cell lines: the EBV-LCL cell lines ratio of 1:0.5:0.5 in hRPMI medium containing 10% FBS and 1% penicillin/streptomycin in presence of 500 U/ml of IL-2, for 5 to 15 days, and during the co-culturing, replacing the hRPMI medium with another hRPMI medium containing 500 U/ml of IL-2 once every 3 to 4 days;
(b) treating the cultured natural killer cells with an HDAC (Histone deacetylase) inhibitor; and
(c) additionally culturing the treated natural killer cells,
wherein the natural killer cells are first treated with the HDAC inhibitor on day 6 of co-culturing, and then additionally treated with the HDAC inhibitor 1 to 3 times at intervals of 2 to 4 days, and
wherein the HDAC inhibitor is Suberoylanilide-hydroxamic acid (SAHA) used at a concentration of 62.5 nM.