# US 2017/0283773 A1 · Methods for improving proliferation and stemness of limbal stem cells

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- Number: US 2017/0283773 A1
- Document ID: US20170283773A1
- Status: Published application
- Kind: A1
- Application number: 15/463,737
- Filed: 2017-03-20
- Published: 2017-10-05
- Earliest claimed: 2016-03-29
- Assignees: Catholic University Industry Academic Cooperation Foundation
- Applicants: Catholic University Industry Academic Cooperation Foundation
- Inventors: So-Hyang CHUNG; Hyun Jung LEE
- Web page: https://patentgrep.com/patent/US20170283773A1

## CPC

- C12N 5/0621: {Eye cells, e.g. cornea, iris pigmented cells (photoreceptors C12N5/062)}
- C12N 2501/415: Wnt; Frizzeled
- C12N 2533/30: Synthetic polymers
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Provided are methods for improving the proliferation and sternness of limbal stem cells (LSCs) by adding a Wnt inhibitor to a medium for limbal explant cultures. When human limbal explants are cultured according to the methods, the Wnt/β-catenin signaling is inhibited, resulting in improvement of LSC proliferation, and improvement in the sternness of LSCs, and thus the LSCs may be obtained in high yield. These effects may lead to a higher success rate of transplantation of a limbal explant outgrowth sheet into a LSCD patient.

## Claims

### Claim 1

A method for improving the proliferation of limbal stem cells, comprising:
(a) immersing a human limbal explant in a medium; and
(b) adding a Wnt inhibitor to the medium.

### Claim 2 (depends on 1)

The method of claim 1, wherein the Wnt inhibitor is N-(6-Methyl-2-benzothiazolyl)-2-[(3,4,6,7-tetrahydro-4-oxo-3-phenylthieno[3,2-d]pyrimidin-2-yl)thio]-acetamide (IWP2).

### Claim 3 (depends on 1)

The method of claim 1, wherein the Wnt inhibitor reduces an amount of β-catenin in cells.

### Claim 4 (depends on 1)

The method of claim 1, wherein the limbal explant is prepared by placing an epithelial part of a limbal tissue face-up on a polyester membrane insert.

### Claim 5

A method for improving the stemness of limbal stem cells, comprising:
(a) immersing a human limbal explant in a medium; and
(b) adding a Wnt inhibitor to the medium.

### Claim 6 (depends on 5)

The method of claim 5, wherein the Wnt inhibitor is N-(6-Methyl-2-benzothiazolyl)-2-[(3,4,6,7-tetrahydro-4-oxo-3-phenylthieno[3,2-d]pyrimidin-2-yl)thio]-acetamide (IWP2).

### Claim 7 (depends on 5)

The method of claim 5, wherein the Wnt inhibitor reduces an amount of β-catenin in cells.

### Claim 8 (depends on 5)

The method of claim 5, wherein the limbal explant is prepared by placing an epithelial part of a limbal tissue face-up on a polyester membrane insert.

### Claim 9 (depends on 5)

The method of claim 5, wherein the method improves a colony forming efficiency (CFE) of the limbal stem cells.