Differentiation of Human Embryonic Stem Cells

• US 20100112692A1
• Filed: 10/22/2009
• Published: 05/06/2010
• Est. Priority Date: 10/31/2008
• Status: Active Grant

• Current Assignee: Janssen Biotech, Inc. (Johnson & Johnson)
• Original Assignee: Janssen Biotech, Inc. (Johnson & Johnson)
• Inventors: Rezania, Alireza
• Granted Patent: US 9,012,218 B2
• Time in Patent Office: Days
• US Class Current: 435/377
• CPC Class Codes: A61P 5/48 of the pancreatic hormones
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/117 Keratinocyte growth factors (KGF-1, i.e. FGF-7; KGF-2, i.e. FGF-12)
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2501/16 Activin; Inhibin; Mullerian inhibiting substance
• CPC Class Codes: C12N 2501/41 Hedgehog proteins; Cyclopamine (inhibitor)
• CPC Class Codes: C12N 2501/415 Wnt; Frizzeled
• CPC Class Codes: C12N 2501/70 Enzymes
• CPC Class Codes: C12N 2501/998 Proteins not provided for elsewhere
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/0676 Pancreatic cells

Abstract

The present invention provides methods to promote the differentiation of pluripotent stem cells. In particular, the present invention provides a method to increase the expression of markers associated with the pancreatic endocrine lineage.

Claims

1. A method of increasing the expression of markers associated with the pancreatic endocrine lineage comprising treating cells expressing markers associated with the pancreatic endocrine lineage with medium comprising a sufficient amount of a TGF-β

receptor agonist to cause an increase in expression of markers associated with the pancreatic endocrine lineage.

2. The method of claim 1, wherein the TGF-β

receptor agonist is selected from the group consisting of activin A, activin B, activin C, peptide variants of activin A, GDF-8, GDF-11 and GDF-15.

3. The method of claim 1, wherein the cells expressing markers associated with the pancreatic endocrine lineage are enriched.

4. A method for differentiating pluripotent stem cells, comprising the steps of:

a. Culturing the pluripotent stem cells,

b. Differentiating the pluripotent stem cells into cells expressing markers characteristic of the definitive endoderm lineage,

c. Differentiating the cells expressing markers characteristic of the definitive endoderm lineage into cells expressing markers characteristic of the pancreatic endoderm lineage, and

d. Differentiating the cells expressing markers characteristic of the pancreatic endoderm lineage into cells expressing markers characteristic of the pancreatic endocrine lineage.

5. The method of claim 4, wherein the expression of markers characteristic of the pancreatic endocrine lineage is increased by treating the cells expressing markers characteristic of the pancreatic endocrine lineage with medium comprising a sufficient amount of a TGF-β

receptor agonist to cause an increase in expression of markers associated with the pancreatic endocrine lineage.

6. The method of claim 4, wherein the TGF-β

receptor agonist is selected from the group consisting of activin A, activin B, activin C, peptide variants of activin A, GDF-8, GDF-11 and GDF-15.

7. The method of claim 4, wherein the cells expressing markers associated with the pancreatic endocrine lineage are enriched.