US 12,351,824 B2
Method for producing natural killer cells from pluripotent stem cells
Yu Qian, Cambridge, MA (US); Hui-Hsin Chang, Cambridge, MA (US); Xi Shi, Cambridge, MA (US); Jianxin Hu, Cambridge, MA (US); and Lan Cao, Cambridge, MA (US)
Assigned to Takeda Pharmaceutical Company Limited, Osaka (JP)
Filed by Takeda Pharmaceutical Company Limited, Osaka (JP)
Filed on Feb. 26, 2021, as Appl. No. 17/186,877.
Claims priority of provisional application 62/983,511, filed on Feb. 28, 2020.
Prior Publication US 2021/0292713 A1, Sep. 23, 2021
Int. Cl. C12N 5/0783 (2010.01); A61K 40/15 (2025.01); A61K 40/31 (2025.01); A61K 40/35 (2025.01); A61K 40/42 (2025.01); C07K 14/54 (2006.01); C07K 14/705 (2006.01); C07K 14/725 (2006.01); C07K 16/28 (2006.01)
CPC C12N 5/0646 (2013.01) [A61K 40/15 (2025.01); A61K 40/31 (2025.01); A61K 40/35 (2025.01); A61K 40/4211 (2025.01); C07K 14/5443 (2013.01); C07K 14/705 (2013.01); C07K 14/7051 (2013.01); C07K 14/70517 (2013.01); C07K 14/70578 (2013.01); C07K 16/2803 (2013.01); A61K 2239/31 (2023.05); A61K 2239/38 (2023.05); A61K 2239/48 (2023.05); C07K 2319/03 (2013.01); C12N 2500/02 (2013.01); C12N 2501/115 (2013.01); C12N 2501/125 (2013.01); C12N 2501/155 (2013.01); C12N 2501/165 (2013.01); C12N 2501/2302 (2013.01); C12N 2501/2307 (2013.01); C12N 2501/26 (2013.01); C12N 2501/515 (2013.01); C12N 2501/998 (2013.01); C12N 2501/999 (2013.01); C12N 2506/45 (2013.01)] 16 Claims
 
1. A method of producing pluripotent stem cell-derived natural killer (NK) cells comprising:
(A) providing a bulk cell population in a bulk cell medium, comprising hematopoietic progenitor cells (HPC) (HP cell bulk) derived from pluripotent stem cells;
(B) culturing HP cell bulk in a CD4/CD8 induction medium comprising a p38 inhibitor and SDF-1 to obtain a heterogeneous population of CD4+/− cells and CD8+/− cells (DP cell bulk);
(C) culturing the DP cell bulk in NK induction medium comprising IL-2 or IL-7 to obtain NK cells; wherein step (C) does not include a step of isolating CD4+/CD8+ cells from the DP cell bulk and wherein >50% of the cells produced from step (C) are NK cells.