# US 10,119,120 B2 · Method for inducing differentiation of pluripotent stem cells into neural precursor cells

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,119,120 B2
- Document ID: US10119120B2
- Status: Granted
- Kind: B2
- Application number: 13/390,225
- Filed: 2010-08-12
- Published: 2012-06-07
- Granted: 2018-11-06
- Earliest claimed: 2009-08-12
- Assignees: Kyoto University
- Applicants: Jun Takahashi; Asuka Morizane
- Inventors: Jun Takahashi; Asuka Morizane
- Web page: https://patentgrep.com/patent/US10119120B2

## CPC

- C12N 5/0618: {Cells of the nervous system}
- C12N 2501/155: Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
- C12N 2506/02: from embryonic cells
- C12N 2506/45: from artificially induced pluripotent stem cells
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention provides a method for inducing differentiation of pluripotent stem cells into neural precursor cells, comprising culturing the pluripotent stem cells in the presence of a small molecule BMP inhibitor, and induced neural precursor cells prepared by this method.

## Claims

### Claim 1

A method for inducing differentiation of a pluripotent cell into a neural precursor cell, the method comprising:
treating the pluripotent stem cell with a p160-Rho-associated coiled-coil kinase (ROCK) inhibitor; and
culturing the treated pluripotent stem cell for inducing the differentiation into the neural precursor cell in a culture medium comprising substances consisting of 4-(6-(4-(piperazin-1-yl)phenyl)pyrazolo[1,5-a]pyrimidin-3-yl)quinoline (LDN-193189) and a TGFβ family inhibitor selected from the group consisting of 4-[4-(1,3-benzodioxol-5-yl)-5-(2-pyridinyl)-1H-imidazol-2-yl]benzamide (SB431542) and 3-(6-methylpyridin-2-yl)-1-phenylthiocarbamoyl-4-quinolin-4-ylpyrazole (A-83-01), wherein the concentration of LDN-193189 is 50 nM or higher.

### Claim 2 (depends on 1)

The method of claim 1, wherein the culture medium further comprises a feeder cell, which is a stromal cell.

### Claim 3 (depends on 2)

The method of claim 2, wherein the stromal cell is a mouse stromal cell.

### Claim 4 (depends on 1)

The method of claim 1, wherein the culture medium is serum free and prior to the culturing of the differentiation induction an embryoid body is formed from the pluripotent stem cell.

### Claim 5 (depends on 1)

The method of claim 1, wherein the culture is performed on a Matrigel coated dish without feeder cells.

### Claim 6 (depends on 1)

The method of claim 1, wherein the pluripotent stem cell is an embryonic stem cell or an induced pluripotent stem cell.

### Claim 7 (depends on 1)

The method of claim 1, wherein the TGFβ family inhibitor is 4-[4-(1,3-benzodioxol-5-yl)-5-(2-pyridinyl)-1H-imidazol-2-yl]benzamide (SB431542).

### Claim 8 (depends on 1)

The method of claim 1, wherein the TGFα family inhibitor is 3-(6-methylpyridin-2-yl)-1-phenylthiocarbamoyl-4-quinolin-4-ylpyrazole(A-83-01).

### Claim 9 (depends on 1)

The method of claim 1, wherein the pluripotent stem cell is an embryonic stem cell.

### Claim 10 (depends on 1)

The method of claim 1, wherein the pluripotent stem cell is an induced pluripotent stem cell.