# US 2021/0340500 A1 · Method for producing intestinal cells from pluripotent stem cells

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- Number: US 2021/0340500 A1
- Document ID: US20210340500A1
- Status: Published application
- Kind: A1
- Application number: 17/274,543
- Filed: 2019-09-09
- Published: 2021-11-04
- Earliest claimed: 2018-09-10
- Assignees: Tokyo Institute of Technology; The University of Tokyo; Kanto Kagaku Kabushiki Kaisha
- Applicants: Tokyo Institute of Technology; The University of Tokyo; Kanto Kagaku Kabushiki Kaisha
- Inventors: Shoen KUME; Nobuaki SHIRAKI; Kazuya MAEDA; Hiroyuki KUSUHARA; Masaya ISHIKAWA; Teruhiko WATANABE
- Web page: https://patentgrep.com/patent/US20210340500A1

## CPC

- C12N 5/0679: {Cells of the gastro-intestinal tract}
- C12N 2500/30: Organic components
- C12N 2500/38: Vitamins
- C12N 2501/12: Hepatocyte growth factor [HGF]
- C12N 2501/16: Activin; Inhibin; Mullerian inhibiting substance
- C12N 2501/39: Steroid hormones
- C12N 2501/727: Kinases (EC 2.7.)
- C12N 2501/999: Small molecules not provided for elsewhere
- C12N 2502/70: Non-animal cells
- C12N 2533/54: Collagen; Gelatin
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## 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.

## Claims

### Claim 1

An enterocyte differentiation medium comprising: 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.

### Claim 2 (depends on 1)

The enterocyte differentiation medium according to claim 1, wherein the GSK3 inhibitor is BIO.

### Claim 3 (depends on 1)

The enterocyte differentiation medium according to claim 1, wherein the activator of a hepatocyte growth factor receptor is HGF.

### Claim 4 (depends on 1)

The enterocyte differentiation medium according to claim 1, wherein the adrenal cortex hormone is dexamethasone.

### Claim 5 (depends on 1)

A method for producing enterocytes, comprising the following steps (1) to (3):
(1) culturing pluripotent stem cells in a medium containing an activator of activin receptor-like kinase-4,7;
(2) culturing the cells resulting from the step (1) in a medium containing a GSK3 inhibitor and a γ-secretase inhibitor; and
(3) culturing the cells resulting from the step (2) on a high density collagen gel membrane in the enterocyte differentiation medium according to claim 1, to obtain enterocytes.

### Claim 6 (depends on 5)

The method for producing enterocytes according to claim 5, wherein the cells resulting from the step (1) are cultured on a high density collagen gel membrane in the step (2).

### Claim 7 (depends on 5)

The method for producing enterocytes according to claim 5, wherein the pluripotent stem cells are cultured on M15 cells in the step (1).

### Claim 8 (depends on 5)

The method for producing enterocytes according to claim 5, wherein the activator of activin receptor-like kinase-4,7 is activin A in the step (1).

### Claim 9 (depends on 5)

The method for producing enterocytes according to claim 5, wherein the GSK3 inhibitor is BIO, and the γ-secretase inhibitor is DAPT in the step (2).

### Claim 10 (depends on 5)

Enterocytes produced by the method according to claim 5.