# US 2014/0193904 A1 · Pdx1 expressing endoderm

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- Number: US 2014/0193904 A1
- Document ID: US20140193904A1
- Status: Published application
- Kind: A1
- Application number: 14/146,510
- Filed: 2014-01-02
- Published: 2014-07-10
- Earliest claimed: 2004-04-27
- Assignees: ViaCyte, Inc.
- Applicants: Viacyte, Inc.
- Inventors: Kevin Allen D'Amour; Alan D. Agulnick; Susan Eliazer; Emmanuel E. Baetge
- Web page: https://patentgrep.com/patent/US20140193904A1

## CPC

- C12N 5/0603: {Embryonic cells (production of embryos, nuclear transfer A01K67/027); Embryoid bodies}
- C12N 5/0606: {Pluripotent embryonic cells, e.g. embryonic stem cells [ES] (embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696)}
- C12N 2501/115: Basic fibroblast growth factor (bFGF, FGF-2)
- C12N 2501/119: Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10
- C12N 2501/15: Transforming growth factor beta (TGF-β)
- C12N 2501/155: Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
- C12N 2501/16: Activin; Inhibin; Mullerian inhibiting substance
- C12N 2501/385: of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR]
- C12N 2501/415: Wnt; Frizzeled
- C12N 2502/13: connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
- C12N 2506/02: from embryonic cells
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Disclosed herein are cell cultures comprising PDX1-positive endoderm cells and methods of producing the same. Also disclosed herein are cell populations comprising substantially purified PDX1-positive endoderm cells as well as methods for enriching, isolating and purifying PDX1-positive endoderm cells from other cell types. Methods of identifying differentiation factors capable of promoting the differentiation of endoderm cells, such as PDX1-positive foregut endoderm cells and PDX1-negative definitive endoderm cells, are also disclosed.

## Claims

### Claim at position 1

-129. (canceled)

### Claim 130

An in vitro cell culture comprising anterior endoderm cells.

### Claim 131 (depends on 130)

The cell culture of claim 130, further comprising a retinoid.

### Claim 132 (depends on 131)

The cell culture of claim 131, wherein the retinoid is retinoic acid (RA).

### Claim 133 (depends on 132)

The cell culture of claim 132, wherein the RA is provided at a concentration greater than 0.2 μM.

### Claim 134 (depends on 132)

The cell culture of claim 132, wherein RA is provided in a concentration ranging from about 0.2 μM to about 50 μM.

### Claim 135 (depends on 132)

The cell culture of claim 132, wherein RA is provided in a concentration of about 1 μM.

### Claim 136 (depends on 130)

The cell culture of claim 130, wherein the anterior endoderm cells are derived from human pluripotent cells.

### Claim 137 (depends on 130)

The cell culture of claim 130, wherein the anterior endoderm cells are derived from definitive endoderm cells.

### Claim 138 (depends on 130)

The cell culture of claim 130, wherein the anterior endoderm cells express HOXa3.

### Claim 139 (depends on 130)

The cell culture of claim 130, wherein the anterior endoderm cells do not express alpha-fetoprotein, CDX1, HOXC6, HOXA13, SOX1 or NFM.

### Claim 140 (depends on 130)

The cell culture of claim 130, wherein the anterior endoderm cells are multipotent cells that can differentiate into the pharynx, lung, esophagus or trachea.

### Claim 141

An in vitro cell population comprising human cells that express HOXa3.

### Claim 142 (depends on 141)

The cell population of claim 141, wherein the human cells express HOXa3 to a greater extent than HOXc6 or HOXa13.

### Claim 143 (depends on 141)

The cell population of claim 141, further comprising a retinoid.

### Claim 144 (depends on 143)

The cell population of claim 143, wherein the retinoid is retinoic acid (RA).

### Claim 145 (depends on 144)

The cell population of claim 144, wherein RA is provided in a concentration of about 1 μM.

### Claim 146 (depends on 141)

The cell population of claim 141, wherein the human cells that express HOXa3 are derived from human pluripotent cells.

### Claim 147

A method of making human anterior endoderm cells in vitro comprising (a) obtaining definitive endoderm cells; and (b) contacting the definitive endoderm cells with a retinoid thereby making anterior endoderm cells in vitro.

### Claim 148 (depends on 147)

The method of claim 147, wherein the anterior endoderm cells express HOXa3.

### Claim 149 (depends on 147)

The method of claim 147, wherein the retinoid is retinoic acid (RA).

### Claim 150 (depends on 149)

The method of claim 149, wherein RA is provided in a concentration of about 1 μM.