# US 2022/0033770 A1 · Macs-based purification of stem cell-derived retinal pigment epithelium

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- Number: US 2022/0033770 A1
- Document ID: US20220033770A1
- Status: Published application
- Kind: A1
- Application number: 17/493,533
- Filed: 2021-10-04
- Published: 2022-02-03
- Earliest claimed: 2015-09-08
- Assignees: FUJIFILM Cellular Dynamics, Inc.
- Applicants: FUJIFILM Cellular Dynamics, Inc.
- Inventors: Nathan MEYER; Lucas CHASE; Casey STANKEWICZ
- Web page: https://patentgrep.com/patent/US20220033770A1

## CPC

- C12N 5/0081: {Purging biological preparations of unwanted cells}
- C12N 5/0621: {Eye cells, e.g. cornea, iris pigmented cells (photoreceptors C12N5/062)}
- C12N 2506/03: from non-embryonic 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

Provided herein are methods of enriching a retinal pigment epithelium (RPE) cell population derived from stem cells. Such a method may comprise removing contaminating cells through the depletion of CD24 positive cells, CD56 positive cells, and/or CD90 positive cells from a starting population of RPE cells.

## Claims

### Claim 1

A method for providing an enriched population of retinal pigment epithelial (RPE) cells comprising:
a) obtaining a starting cell population comprising RPE cells; and
b) enriching said starting cell population for RPE cells by removing therefrom cells that are positive for CD24, cells that are positive for CD56 and/or cells that are positive for CD90, thereby providing a RPE-enriched cell population that is enriched for RPE cells as compared to the starting cell population.

### Claim 2 (depends on 1)

The method of claim 1, further comprising determining a level of enrichment of the RPE cells in said RPE-enriched population.

### Claim 3 (depends on 2)

The method of claim 2, wherein the level of enrichment is determined through the use of a retinal epithelial-specific marker selected from the group consisting of BEST1, CRALBP, TYRP1, PMEL17, and MITF.

### Claim 4 (depends on 1)

The method of claim 1, wherein the RPE-enriched cell population is enriched for RPE cells as compared to the starting cell population as determined by BEST1 sorting.

### Claim 5 (depends on 1)

The method of claim 1, wherein the RPE-enriched population is at least 95% RPE cells.

### Claim 6 (depends on 5)

The method of claim 5, wherein the RPE-enriched population is at least 99% RPE cells.

### Claim 7 (depends on 6)

The method of claim 6, wherein the RPE-enriched population is essentially pure RPE cells.

### Claim 8 (depends on 1)

The method of claim 1, wherein the RPE cells are human RPE cells.

### Claim 9 (depends on 1)

The method of claim 1, wherein the starting cell population is prepared from pluripotent stem cells.

### Claim 10 (depends on 9)

The method of claim 9, wherein the pluripotent stem cells are induced pluripotent stem cells.

### Claim 11 (depends on 1)

The method of claim 1, wherein the cells that are positive for CD24, cells that are positive for CD56 and/or cells that are positive for CD90 are removed by magnetic bead-based sorting or fluorescence-based sorting.

### Claim 12 (depends on 1)

The method of claim 1, wherein the cells that are positive for CD24, cells that are positive for CD56 and/or cells that are positive for CD90 are removed using an antibody or an aptamer that recognizes CD24, CD56 and/or CD90.

### Claim 13 (depends on 1)

The method of claim 1, wherein the RPE-enriched cell population is not genetically modified.

### Claim 14 (depends on 1)

The method of claim 1, wherein step b) is carried out without genetically engineering the starting cell population.

### Claim 15 (depends on 1)

The method of claim 1, wherein said starting cell population is enriched for RPE cells by removing therefrom cells that are positive for CD24.

### Claim 16 (depends on 1)

The method of claim 1, wherein said starting cell population is enriched for RPE cells by removing therefrom cells that are positive for CD56.

### Claim 17 (depends on 1)

The method of claim 1, wherein said starting cell population is enriched for RPE cells by removing therefrom cells that are positive for CD90.

### Claim 18 (depends on 1)

The method of claim 1, wherein said starting cell population is enriched for RPE cells by removing therefrom cells that are positive for CD90, cells that are positive of CD56 and cells that are positive for CD24.