# US 10,370,645 B2 · Endocrine precursor cells, pancreatic hormone-expressing cells and methods of production

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- Number: US 10,370,645 B2
- Document ID: US10370645B2
- Status: Granted
- Kind: B2
- Application number: 15/173,316
- Filed: 2016-06-03
- Published: 2016-12-08
- Granted: 2019-08-06
- Earliest claimed: 2006-03-02
- Assignees: Emory University; Dana-Farber Cancer Institute, Inc.; Brigham and Women's Hopsital; President and Fellows of Harvard College
- Applicants: ViaCyte, Inc.
- Inventors: Kevin A. D'Amour; Anne Bang; Emmanuel E. Baetge
- Web page: https://patentgrep.com/patent/US10370645B2

## CPC

- C12N 5/0676: {Pancreatic cells}
- A61K 35/39: Pancreas; Islets of Langerhans (Langerhans cells of epidermis A61K35/36)
- A61P 1/18: for pancreatic disorders, e.g. pancreatic enzymes
- A61P 3/10: for hyperglycaemia, e.g. antidiabetics
- A61P 5/48: of the pancreatic hormones
- C12N 5/0678: {Stem cells; Progenitor cells; Precursor cells}
- C12N 2500/38: Vitamins
- C12N 2501/10: Growth factors
- C12N 2501/105: Insulin-like growth factors [IGF]
- 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 2501/12: Hepatocyte growth factor [HGF]
- C12N 2501/15: Transforming growth factor beta (TGF-β)
- C12N 2501/16: Activin; Inhibin; Mullerian inhibiting substance
- C12N 2501/33: Insulin
- C12N 2501/335: Glucagon; Glucagon-like peptide [GLP]; Exendin
- C12N 2501/385: of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR]
- C12N 2501/41: Hedgehog proteins; Cyclopamine (inhibitor)
- C12N 2501/415: Wnt; Frizzeled
- C12N 2501/42: Notch; Delta; Jagged; Serrate
- C12N 2501/58: Adhesion molecules, e.g. ICAM, VCAM, CD18 (ligand), CD11 (ligand), CD49 (ligand)
- C12N 2501/734: Proteases (EC 3.4.)
- C12N 2501/999: Small molecules not provided for elsewhere
- 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)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

Disclosed herein are cell cultures and enriched cell populations of endocrine precursor cells, immature pancreatic hormone-expressing cells and mature pancreatic hormone-expressing cells. Also disclosed herein are methods of producing such cell cultures and cell populations.

## Claims

### Claim 1

An in vitro method of making a population of human pancreatic and duodenal homeobox (PDX1)-positive pancreatic endoderm, comprising:
a) obtaining a population of human definitive endoderm cells; and
b) contacting the population of human definitive endoderm cells from step (a) with a retinoid, thereby making the population of human PDX1-positive pancreatic endoderm.

### Claim 2 (depends on 1)

The method of claim 1, further comprising contacting the population of human definitive endoderm cells from step (a) with the gamma secretase inhibitor.

### Claim 3 (depends on 2)

The method of claim 2, wherein the gamma secretase inhibitor is N—[N-(3,5-Diflurophenaacetyl-L-alanyl)]-S-phenylglycine t-Butyl Ester (DAPT).

### Claim 4 (depends on 1)

The method of claim 1, further comprising transplanting the population of human PDX1-positive endoderm cells into a human host to mature into insulin secreting cells.

### Claim 5 (depends on 4)

The method of claim 4, wherein the insulin secreting cells form islet-like cell clusters.

### Claim 6 (depends on 5)

The method of claim 5, wherein the islet-like cell clusters comprise singly-positive insulin secreting cells.

### Claim 7 (depends on 1)

The method of claim 1, wherein the population of human PDX1-positive pancreatic endoderm cells expresses neurogenin (NGN3) and does not substantially express a marker selected from the group consisting of ghrelin, glucagon, insulin, and islet amyloid polypeptide (IAPP).

### Claim 8 (depends on 1)

The method of claim 1, further comprising contacting the population of human PDX1-positive pancreatic endoderm cells with a factor selected from the group consisting of nicotinamide (NIC), exendin 4 (Ex4), hepatocyte growth factor (HGF), insulin-like growth factor (IGF) and combinations thereof.

### Claim 9 (depends on 1)

The method of claim 1, wherein the retinoid is retinoic acid.