# US 10,457,913 B2 · Corneal epithelioid cells derived from surface ectodermal cells

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- Number: US 10,457,913 B2
- Document ID: US10457913B2
- Status: Granted
- Kind: B2
- Application number: 15/543,021
- Filed: 2016-01-12
- Published: 2018-05-31
- Granted: 2019-10-29
- Earliest claimed: 2015-01-13
- Assignees: Osaka University
- Applicants: Osaka University
- Inventors: Kohji Nishida; Yuzuru Sasamoto; Ryuhei Hayashi
- Web page: https://patentgrep.com/patent/US10457913B2

## CPC

- C12N 5/0602: {Vertebrate cells}
- C12N 5/0621: {Eye cells, e.g. cornea, iris pigmented cells (photoreceptors C12N5/062)}
- C07K 14/4702: {Regulators; Modulating activity}
- C12N 5/10: Cells modified by introduction of foreign genetic material
- C12N 15/86: Viral vectors
- C12N 2501/11: Epidermal growth factor [EGF]
- C12N 2501/60: Transcription factors
- C12N 2501/603: Oct-3/4
- C12N 2501/604: Klf-4
- C12N 2506/097: from oral mucosa cells
- C12N 2510/00: Genetically modified cells
- C12N 2740/15043: viral genome or elements thereof as genetic vector
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)

## Abstract

The present invention relates to a method for inducing a keratin 12-positive corneal epithelioid cell from a surface ectoderm-derived cell. More specifically, the present invention relates to a method for inducing a keratin 12-positive corneal epithelioid cell, comprising introducing PAX6, KLF4, and OCT4 into a surface ectoderm-derived cell, such as an oral mucosal epithelial cell, and a corneal epithelioid cell induced by the method.

## Claims

### Claim 1

An in vitro or ex vivo method for producing a keratin-12 positive human corneal epithelioid cell, comprising:
introducing a lentiviral vector comprising nucleic acids encoding PAX6, KLF4, and OCT4 into a human epithelial cell,
wherein expression of the lentiviral vector induces the production of a keratin 12-positive human corneal epithelioid cell.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the human corneal epithelioid cell is induced not through a state of a pluripotent cell.

### Claim 3 (depends on 1)

The method according to claim 1, wherein the human epithelial cell is an oral mucosal epithelial cell.

### Claim 4 (depends on 1)

A method for producing a human corneal epithelioid cell sheet, comprising producing human corneal epithelioid cells by the method according to claim 1 and stratifying the cells.

### Claim 5 (depends on 1)

The method according to claim 1, wherein the lentiviral vector further comprises nucleic acids encoding two PAX6 isoforms, Pa and Pb.

### Claim 6 (depends on 5)

The method according to claim 5, wherein the corneal epithelioid cell is further keratin 3-positive.

### Claim 7 (depends on 1)

The method according to claim 1, the human epithelial cell is selected from the group consisting of epidermal cell, oral mucosal epithelial cell, and corneal epithelial cell.