公开(公告)号：US20170081639A1

公开(公告)日：2017-03-23

申请号：US15312273

申请日：2015-05-19

Applicant: TOKYO INSTITUTE OF TECHNOLOGY

Inventor： Shoen KUME ,  Nobuaki SHIRAKI

IPC: C12N5/071 ,  G01N33/50

CPC classification number: C12N5/0676 ,  A61K35/12 ,  C12N5/0678 ,  C12N15/09 ,  C12N2500/92 ,  C12N2500/98 ,  C12N2501/01 ,  C12N2501/119 ,  C12N2501/15 ,  C12N2501/155 ,  C12N2501/16 ,  C12N2501/385 ,  C12N2501/41 ,  C12N2501/415 ,  C12N2501/727 ,  C12N2501/73 ,  C12N2506/45 ,  C12N2533/52 ,  G01N33/507 ,  G01N33/5073 ,  G01N33/74

Abstract: It is an object of the present invention to provide a method for efficiently directing differentiation into insulin-producing cells in a xeno-free culture system. According to the present invention, there is provided a method for directed differentiation into insulin-producing cells, comprising culturing stem cells in the following steps (1) to (5): (1) a step of culturing stem cells in a medium comprising an activator of activin receptor-like kinase-4/-7 and a GSK3 inhibitor and then culturing in a medium comprising an activator of activin receptor-like kinase-4/-7; (2) a step of culturing the cells obtained in step (1) in a medium comprising a hedgehog signaling inhibitor and an FGF; (3) a step of culturing the cells obtained in step (2) in a medium comprising a retinoic acid receptor agonist, a hedgehog signaling inhibitor and a BMP signaling inhibitor; (4) a step of culturing the cells obtained in step (3) in a medium comprising a TGF-β type I activin receptor-like kinase-4/-5/-7 inhibitor and a BMP signaling inhibitor; and (5) a step of culturing the cells obtained in step (4) in a medium comprising a phosphodiesterase inhibitor.

公开(公告)号：US20210115397A1

公开(公告)日：2021-04-22

申请号：US17050312

申请日：2019-04-25

Applicant: TOKYO INSTITUTE OF TECHNOLOGY ,  Ajinomoto Co., Inc.

Inventor： Shoen KUME ,  Nobuaki SHIRAKI ,  Akira CHIBA ,  Takayuki ENOMOTO

IPC: C12N5/074

Abstract: In order to improve the efficiency of inducing differentiation of pluripotent stem cells, provided is a method for promoting differentiation of pluripotent stem cells, the method including a step of culturing pluripotent stem cells in a medium, wherein the medium is a medium containing a) an insulin-like growth factor, b) an insulin analogue preparation containing no zinc, c) an insulin analogue preparation containing a low concentration of zinc, or d) a compound exhibiting an insulin-like action.

公开(公告)号：US20210340500A1

公开(公告)日：2021-11-04

申请号：US17274543

申请日：2019-09-09

Applicant: TOKYO INSTITUTE OF TECHNOLOGY ,  THE UNIVERSITY OF TOKYO ,  KANTO KAGAKU KABUSHIKI KAISHA

Inventor： Shoen KUME ,  Nobuaki SHIRAKI ,  Kazuya MAEDA ,  Hiroyuki KUSUHARA ,  Masaya ISHIKAWA ,  Teruhiko WATANABE

IPC: C12N5/071

Abstract: As a method for producing enterocytes derived from pluripotent stem cells and having functions corresponding to those of actual enterocytes, a method for producing enterocytes using an enterocyte differentiation medium containing a GSK3 inhibitor, and at least one selected from the group consisting of an activator of a hepatocyte growth factor receptor, an adrenal cortex hormone, calcitriol, and dimethyl sulfoxide is provided.

公开(公告)号：US20210207085A1

公开(公告)日：2021-07-08

申请号：US16755490

申请日：2018-10-09

Applicant: TOKYO INSTITUTE OF TECHNOLOGY ,  KANTO KAGAKU KABUSHIKI KAISHA ,  CHUGAI SEIYAKU KABUSHIKI KAISHA

Inventor： Shoen KUME ,  Nobuaki SHIRAKI ,  Hiroyuki YAMAGUCHI ,  Tomoaki INOUE ,  Toshito NAKAGAWA ,  Jumpei KIYOKAWA

IPC: C12N5/071

Abstract: The present invention provides, in order to prepare matured hepatocytes analogous in various points to primary hepatocytes, a method for preparing hepatocytes or cells that can be differentiated into hepatocytes from pluripotent stem cells, comprising the steps of: (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), cells are cultured on a high-density collagen gel membrane.