Patent ID: 11859212
Assignee: PUBLIC UNIVERSITY CORPORATION NAGOYA CITY UNIVERSITY
Field: Measurement (Instruments)
Classification: CPC C  A  G | IPC A  C  G

Claim 0:
1. A method for preparing an intestinal organoid from pluripotent stem cells, comprising the following steps (1) to (4):
(1) differentiating pluripotent stem cells into endoderm-like cells;
(2) differentiating the endoderm-like cells obtained in step (1) into intestinal stem cell-like cells;
(3) culturing the intestinal stem cell-like cells obtained in step (2) to form spheroids; and
(4) differentiating the spheroids formed in step (3) to form an intestinal organoid, the step including culture in the presence of a MEK1/2 inhibitor, a DNA methylation inhibitor, a TGF-β receptor inhibitor, and a γ-secretase inhibitor, in addition to an epidermal growth factor, a BMP inhibitor, and a Wnt signal activator,
wherein step (4) includes the following steps (4-1) and (4-2):
(4-1) culture in the presence of the epidermal growth factor, the BMP inhibitor, and the Wnt signal activator; and
(4-2) culture in the presence of the MEK1/2 inhibitor, the DNA methylation inhibitor, the TGF-β receptor inhibitor, and the γ-secretase inhibitor, in addition to the epidermal growth factor, the BMP inhibitor, and the Wnt signal activator;
wherein step (4-1) precedes step (4-2).