# US 2014/0193905 A1 · Expansion of definitive endoderm cells

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 2014/0193905 A1
- Document ID: US20140193905A1
- Status: Published application
- Kind: A1
- Application number: 14/155,963
- Filed: 2014-01-15
- Published: 2014-07-10
- Earliest claimed: 2003-12-23
- Assignees: ViaCyte, Inc.
- Applicants: ViaCyte, Inc.
- Inventors: Olivia Kelly; Emmanuel E. Baetge; Melissa Carpenter
- Web page: https://patentgrep.com/patent/US20140193905A1

## 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 2500/90: Serum-free medium, which may still contain naturally-sourced components
- C12N 2501/115: Basic fibroblast growth factor (bFGF, FGF-2)
- 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 expanded definitive endoderm cells as well as methods for expanding definitive endoderm cells in culture.

## Claims

### Claim at position 1

-11. (canceled)

### Claim 12

An in vitro population of passaged human definitive endoderm cells.

### Claim 13 (depends on 12)

The cell population of claim 12, wherein the population of passaged human definitive endoderm cells expresses SOX17, GSC, MIXL1 or CXCR4.

### Claim 14 (depends on 12)

The cell population of claim 12, wherein the population of passaged human definitive endoderm cells expresses SOX17 and CXCR4.

### Claim 15 (depends on 12)

The cell population of claim 12, wherein the population of passaged human definitive endoderm cells does not express Oct4, Brachyury, ZIC1 or SOX1.

### Claim 16 (depends on 12)

The cell population of claim 12, wherein the population of passaged human definitive endoderm cells is a population of cells that has been passaged at least twice.

### Claim 17 (depends on 12)

The cell population of claim 12, wherein the population of passaged human definitive endoderm cells is in contact with a surface coated with human fibronectin.

### Claim 18 (depends on 12)

The cell population of claim 12, wherein the population of passaged human definitive endoderm cells is in a medium comprising less than 2% (v/v) serum.

### Claim 19 (depends on 12)

The cell population of claim 12, wherein the population of passaged human definitive endoderm cells is in a medium comprising at least one growth factor from the TGFβ superfamily or a member of the epidermal growth factor family.

### Claim 20 (depends on 19)

The cell population of claim 19, wherein the at least one growth factor from the TGFβ superfamily comprises activin A.

### Claim 21 (depends on 12)

The cell population of claim 12, wherein the population of passaged human definitive endoderm cells has been passaged using a protease or by disrupting contacts between cells.

### Claim 22 (depends on 21)

The cell population of claim 21, wherein the protease is trypsin.

### Claim 23 (depends on 12)

The cell population of claim 12, wherein the population of passaged human definitive endoderm cells is derived from human embryonic stem cells (hESCs).

### Claim 24 (depends on 12)

The cell population of claim 12, wherein the population of passaged human definitive endoderm cells is substantially free of pluripotent cells.

### Claim 25 (depends on 24)

The cell population of claim 24, wherein at least 1 passaged human definitive endoderm cell is present for every 5 pluripotent cells.

### Claim 26

A method of making passaged human definitive endoderm cells in vitro comprising passaging definitive endoderm cells thereby forming passaged human definitive endoderm cells.

### Claim 27 (depends on 26)

The method of claim 26, wherein the passaged human definitive endoderm cells express SOX17, GSC, MIXL1 or CXCR4.

### Claim 28 (depends on 26)

The method of claim 26, wherein the passaged human definitive endoderm cells do not express Oct4, Brachyury, ZIC1 or SOX1.

### Claim 29 (depends on 26)

The method of claim 26, wherein the definitive endoderm cells are passaged at least twice.

### Claim 30 (depends on 26)

The method of claim 26, wherein the definitive endoderm cells are derived from human embryonic stem cells (hESCs).

### Claim 31 (depends on 26)

The method of claim 26, wherein 1 passaged human definitive endoderm cell is present for every 5 pluripotent cell.

### Claim 32

A method of expanding human definitive endoderm cells in vitro comprising passaging definitive endoderm cells thereby expanding human definitive endoderm cells.