# US 2016/0257931 A1 · Method for culture of corneal endothelial cells, process for production of corneal endothelial cell sheet for transplantation purposes, and culture kit for corneal endothelial cells

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- Number: US 2016/0257931 A1
- Document ID: US20160257931A1
- Status: Published application
- Kind: A1
- Application number: 15/157,900
- Filed: 2016-05-18
- Published: 2016-09-08
- Earliest claimed: 2010-02-05
- Assignees: Cornea Regeneration Institute Co., Ltd.
- Applicants: Cornea Regeneration Institute Co., Ltd.
- Inventors: Nobuyuki SHIMA; Miwa YAMAUCHI; Masahiro YAMAGUCHI; Satoru YAMAGAMI
- Web page: https://patentgrep.com/patent/US20160257931A1

## CPC

- C12N 5/0621: {Eye cells, e.g. cornea, iris pigmented cells (photoreceptors C12N5/062)}
- C12N 2500/38: Vitamins
- C12N 2501/115: Basic fibroblast growth factor (bFGF, FGF-2)
- 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

The invention provides a method of culturing a corneal endothelial cell by use of a culture medium containing an ascorbic acid derivative and the like.

## Claims

### Claim 1

A method of proliferating a corneal endothelial cell, comprising culturing a corneal endothelial cell in a culture medium containing ascorbyl 2-phosphate, thereby proliferating a corneal endothelial cell.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the corneal endothelial cell is cultured on a biopolymer.

### Claim 3 (depends on 2)

The method according to claim 2, wherein the biopolymer is an extracellular matrix molecule containing collagen.

### Claim 4 (depends on 3)

The method according to claim 3, wherein the collagen is atelocollagen.

### Claim 5 (depends on 1)

A corneal endothelial cell produced by the method according to claim 1.

### Claim 6

A method of producing a corneal endothelial cell sheet for transplantation, comprising a step of culturing a corneal endothelial cell in a culture medium containing ascorbyl 2-phosphate, thereby producing a corneal endothelial cell sheet for transplantation.

### Claim 7 (depends on 6)

The method according to claim 6, wherein the corneal endothelial cell is cultured on a biopolymer.

### Claim 8 (depends on 7)

The method according to claim 7, wherein the biopolymer is an extracellular matrix molecule containing collagen.

### Claim 9 (depends on 8)

The method according to claim 8, wherein the collagen is atelocollagen.

### Claim 10 (depends on 6)

A corneal endothelial cell sheet for transplantation, which is produced by the method according to claim 6.

### Claim 11

A corneal endothelial cell culture kit comprising a substrate coated with a biopolymer and a culture medium containing an ascorbic acid derivative.