Methods of producing pancreatic hormones

• US 9,585,917 B2
• Filed: 08/08/2011
• Issued: 03/07/2017
• Est. Priority Date: 03/02/2006
• Status: Active Grant

• Current Assignee: Viacyte Incorporated (Vertex Pharmaceuticals Incorporated)
• Original Assignee: Viacyte Incorporated (Vertex Pharmaceuticals Incorporated)
• Inventors: Martinson, Laura, Kroon, Evert, D'Amour, Kevin, Baetge, Emmanuel Edward
• Primary Examiner(s): Singh, Anoop
• Assistant Examiner(s): Montanari, David A
• Application Number:
  US13/205,511
• Publication Number:
  US 20120009675A1
• Time in Patent Office: 2,038 Days
• Field of Search: 435/377
• US Class Current: 1/1
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: A61K 38/26 Glucagons
• CPC Class Codes: A61K 38/28 Insulins
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2501/10 Growth factors
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/113 Acidic fibroblast growth factor (aFGF, FGF-1)
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/119 Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10
• CPC Class Codes: C12N 2501/12 Hepatocyte growth factor [HGF]
• CPC Class Codes: C12N 2501/16 Activin; Inhibin; Mullerian inhibiting substance
• CPC Class Codes: C12N 2501/33 Insulin together with transferrin C12N2500/25; Insulin-like growth factors C12N2501/105
• CPC Class Codes: C12N 2501/385 of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR]
• CPC Class Codes: C12N 2501/41 Hedgehog proteins; Cyclopamine (inhibitor)
• CPC Class Codes: C12N 2501/415 Wnt; Frizzeled
• CPC Class Codes: C12N 2501/42 Notch; Delta; Jagged; Serrate
• CPC Class Codes: C12N 2501/58 Adhesion molecules, e.g. ICAM, VCAM, CD18 (ligand), CD11 (ligand), CD49 (ligand)
• CPC Class Codes: C12N 2501/999 Small molecules not provided for elsewhere
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/0676 : Pancreatic cells
• CPC Class Codes: C12N 5/0678 : Stem cells; Progenitor cells; Precursor cells

Abstract

Disclosed herein are methods of producing pancreatic hormone-expressing cells by first differentiating pluripotent cells in cell culture so as to produce endodermal cells, the endodermal cells being competent to further differentiate into hormone-expressing cells capable of secreting at least one pancreatic hormone in response to a physiological signal, and then, transplanting the cultured endodermal cells into an organism, such as an organism in need of an endocrine cell therapy.

Claims

1. A method for lowering blood glucose levels in a mammal, comprising the steps of:

(a) obtaining a population of human definitive endoderm cells;

(b) differentiating the population of definitive endoderm cells into a population of pancreatic endoderm and endocrine precursor cells using a medium comprising glucose, wherein the population expresses NKX6.1 and PDX1;

(c) transplanting the population of pancreatic endoderm and endocrine precursor cells into the mammal; and

(d) maturing the transplanted cells of step (c) into insulin secreting cells.

2. A method for lowering blood glucose levels in a mammal, comprising the steps of:

(a) obtaining a population of human definitive endoderm cells;

(b) differentiating the population of definitive endoderm cells into a population of pancreatic endoderm cells using a DMEM-high glucose medium;

(c) differentiating the population of pancreatic endoderm cells into a population of pancreatic endocrine precursor cells expressing NKX6.1 and PDX1, but not CDX2 using the DMEM-high glucose medium;

(d) transplanting the population of pancreatic endocrine precursor cells into the mammal; and

(e) maturing the transplanted cells of step (d) into insulin secreting cells.

3. A method for lowering blood glucose levels in a mammal, comprising the steps of:

(a) obtaining a population of human definitive endoderm cells;

(b) differentiating the population of definitive endoderm cells into a population of pancreatic endoderm cells using a DMEM-high glucose medium;

(c) differentiating the population of pancreatic endoderm cells into a population of pancreatic endocrine precursor cells expressing NKX6.1 and PDX 1, but not CDX2 using the DMEM-high glucose medium;

(d) differentiating the population of pancreatic endocrine precursor cells into a population of cells expressing insulin using the DMEM-high glucose medium; and

(e) transplanting into the mammal the population of insulin expressing cells.