# US 10,246,679 B2 · Method for producing pluripotent stem cell having improved chimera forming ability from primed pluripotent stem cell, and composition and combination therefor

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- Number: US 10,246,679 B2
- Document ID: US10246679B2
- Status: Granted
- Kind: B2
- Application number: 14/635,331
- Filed: 2015-03-02
- Published: 2016-09-08
- Granted: 2019-04-02
- Earliest claimed: 2015-03-02
- Assignees: The University of Tokyo
- Applicants: The University of Tokyo
- Inventors: Hiromitsu Nakauchi; Hideki Masaki; Hideyuki Murayama
- Web page: https://patentgrep.com/patent/US10246679B2

## CPC

- C12N 5/0606: {Pluripotent embryonic cells, e.g. embryonic stem cells [ES] (embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696)}
- A01K 67/0271: Chimeric vertebrates, e.g. comprising exogenous cells
- C12N 15/85: for animal cells
- A01K 2207/12: Animals modified by administration of exogenous cells
- C12N 2501/235: Leukemia inhibitory factor [LIF]
- C12N 2501/415: Wnt; Frizzeled
- C12N 2501/58: Adhesion molecules, e.g. ICAM, VCAM, CD18 (ligand), CD11 (ligand), CD49 (ligand)
- C12N 2501/60: Transcription factors
- C12N 2510/00: Genetically modified cells
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A01K: ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS

## Abstract

The present invention provides a method for putting a primed pluripotent stem cell in a less differentiated state, a method for producing a pluripotent stem cell having an improved chimera forming ability from a primed pluripotent stem cell, and a method for producing a chimeric animal by using the pluripotent stem cell obtained by any of these methods. The present invention also provides a composition containing a Wnt/β catenin signal inhibitor.

## Claims

### Claim 1

A method of increasing chimera forming ability of a primed pluripotent stem cell, comprising treating a primed pluripotent stem cell with a Wnt/β-catenin signal transduction selective inhibitor and a signal transducer and activator of transcription 3 (STAT3) activator to place the primed pluripotent stem cell in a state where a Wnt/β-catenin signal is selectively inhibited, among Wnt signaling pathways, and a STAT3 signal is activated, wherein following the treatment the primed pluripotent stem cell has increased chimera forming ability compared to an untreated control primed pluripotent stem cell.

### Claim 2 (depends on 1)

A method for producing a chimeric non-human mammal, comprising introducing the pluripotent stem cell produced by the method according to claim 1 into a non-human mammalian blastocyst and developing the blastocyst to produce a chimeric non-human mammal.

### Claim 3 (depends on 1)

The method according to claim 1, wherein the treating comprises treating the primed pluripotent stem cell with a β-catenin degradation accelerator and LIF.

### Claim 4 (depends on 1)

The method according to claim 1, wherein the Wnt/β-catenin signal transduction selective inhibitor comprises XAV939.