US 11,692,173 B2
Method for inducing differentiation of pluripotent stem cells into hepatocytes
Shoen Kume, Tokyo (JP); Nobuaki Shiraki, Tokyo (JP); Hiroyuki Yamaguchi, Isehara (JP); Tomoaki Inoue, Gotemba (JP); Toshito Nakagawa, Gotemba (JP); and Jumpei Kiyokawa, Gotemba (JP)
Assigned to TOKYO INSTITUTE OF TECHNOLOGY, Tokyo (JP); KANTO KAGAKU KABUSHIKI KAISHA, Tokyo (JP); and CHUGAI SEIYAKU KABUSHIKI KAISHA, Tokyo (JP)
Appl. No. 16/755,490
Filed by TOKYO INSTITUTE OF TECHNOLOGY, Tokyo (JP); KANTO KAGAKU KABUSHIKI KAISHA, Tokyo (JP); and CHUGAI SEIYAKU KABUSHIKI KAISHA, Tokyo (JP)
PCT Filed Oct. 9, 2018, PCT No. PCT/JP2018/037531
§ 371(c)(1), (2) Date Apr. 10, 2020,
PCT Pub. No. WO2019/073951, PCT Pub. Date Apr. 18, 2019.
Claims priority of application No. 2017-198201 (JP), filed on Oct. 12, 2017.
Prior Publication US 2021/0207085 A1, Jul. 8, 2021
Int. Cl. C12N 5/071 (2010.01)
CPC C12N 5/067 (2013.01) [C12N 2501/119 (2013.01); C12N 2501/12 (2013.01); C12N 2501/155 (2013.01); C12N 2501/16 (2013.01); C12N 2501/237 (2013.01); C12N 2501/727 (2013.01); C12N 2533/54 (2013.01); C12Y 207/11 (2013.01)] 6 Claims
OG exemplary drawing
 
1. A method for preparing hepatocytes or cells that can be differentiated into hepatocytes from pluripotent stem cells, comprising the following steps (1) to (4):
(1) culturing the pluripotent stem cells in a medium containing an activator of an activin receptor-like kinase-4,7;
(2) culturing the cells obtained in the step (1) in a medium containing a bone morphogenetic factor and a fibroblast growth factor;
(3) culturing the cells obtained in the step (2) in a medium containing an activator of a hepatocyte growth factor receptor and an activator of an oncostatin M receptor; and
(4) culturing the cells obtained in the step (3) to obtain hepatocytes or cells that can be differentiated into hepatocytes, wherein
in at least one of the steps (2), (3) and (4), the cells are cultured on a high-density collagen gel membrane.