Islet cells from human embryonic stem cells

• US 7,033,831 B2
• Filed: 12/06/2002
• Issued: 04/25/2006
• Est. Priority Date: 12/07/2001
• Status: Expired due to Term

• Current Assignee: Asterias Biotherapeutics Incorporated (Lineage Cell Therapeutics, Inc.)
• Original Assignee: Geron Corporation
• Inventors: Fisk, Gregory J., Inokuma, Margaret S.
• Primary Examiner(s): Crouch, Deborah
• Application Number:
  US10/313,739
• Publication Number:
  US 20030138948A1
• Time in Patent Office: 1,236 Days
• Field of Search: 435/363, 435/366, 435/377, 435/69.4, 435/375
• US Class Current: 435/377
• CPC Class Codes: A01N 2300/00 Combinations or mixtures of active ingredients covered by classes A01N27/00 - A01N65/48 with other active or formulation relevant ingredients, e.g. specific carrier materials or surfactants, covered by classes A01N25/00 - A01N65/48
• CPC Class Codes: A01N 65/00 Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof containing compounds of determined constitution A01N27/00 - A01N59/00
• 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: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/50 for increasing or potentiating the activity of insulin
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/30 Organic components metal chelators C12N2500/10; calcitonin C12N2500/14; transferrin C12N2500/24
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2500/44 Thiols, e.g. mercaptoethanol
• CPC Class Codes: C12N 2500/46 Amines, e.g. putrescine
• CPC Class Codes: C12N 2500/62 DMSO
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/12 Hepatocyte growth factor [HGF]
• CPC Class Codes: C12N 2501/13 Nerve growth factor [NGF]; Brain-derived neurotrophic factor [BDNF]; Cilliary neurotrophic factor [CNTF]; Glial-derived neurotrophic factor [GDNF]; Neurotrophins [NT]; Neuregulins
• CPC Class Codes: C12N 2501/148 : Transforming growth factor alpha [TGF-a]
• 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 2501/19 : Growth and differentiation factors [GDF]
• 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 2501/392 : Sexual steroids
• CPC Class Codes: C12N 2501/395 : Thyroid hormones
• CPC Class Codes: C12N 2501/41 : Hedgehog proteins; Cyclopamine (inhibitor)
• CPC Class Codes: C12N 2502/02 : embryonic cells
• CPC Class Codes: C12N 2502/14 : hepatocytes
• CPC Class Codes: C12N 2503/00 : Use of cells in diagnostics
• CPC Class Codes: C12N 2503/02 : Drug screening
• CPC Class Codes: C12N 2506/02 : from embryonic cells
• CPC Class Codes: C12N 2510/00 : Genetically modified cells
• CPC Class Codes: C12N 5/0603 : Embryonic cells production of embryos, nuclear transfer A01K67/027; Embryoid bodies
• CPC Class Codes: C12N 5/0606 : Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0607 : Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0618 : Cells of the nervous system
• CPC Class Codes: C12N 5/067 : Hepatocytes
• CPC Class Codes: C12N 5/0676 : Pancreatic cells
• CPC Class Codes: C12N 5/0678 : Stem cells; Progenitor cells; Precursor cells
• CPC Class Codes: G01N 33/00 : Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
• CPC Class Codes: G01N 33/5008 : for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/5014 : for testing toxicity
• CPC Class Codes: G01N 33/5067 : Liver cells
• CPC Class Codes: G01N 33/507 : Pancreatic cells
• CPC Class Codes: G01N 33/5073 : Stem cells
• CPC Class Codes: G01N 33/56966 : Animal cells

Abstract

This disclosure provides a system for producing pancreatic islet cells from embryonic stem cells. Differentiation is initiated towards endoderm cells, and focused using reagents that promote emergence of islet precursors and mature insulin-secreting cells. High quality populations of islet cells can be produced in commercial quantities for use in research, drug screening, or regenerative medicine.

Claims

1. A method for obtaining human insulin secreting cells, comprising

a) culturing hES cells in a first medium comprising Activin A;

and

b) maturing the cells from a) in a second medium comprising nicotinamide;

thereby obtaining a population comprising human insulin secreting cells.

2. The method of claim 1, wherein the first medium further comprises n-butyrate.

3. A method for obtaining human insulin secreting cells, comprising

a) culturing hES cells in a suspension culture to form embryoid bodies;

b) culturing the cells from a) in a medium comprising trans-retinoic acid;

c) culturing the cells from b) in a medium comprising a TGF-β

antagonist and one or more mitogens; and

d) maturing the cells from c) in a medium comprising nicotinamide;

thereby obtaining a population comprising human insulin-secreting cells.

4. The method of claim 3, wherein the TGF-β

antagonist is Noggin.

5. The method of claim 3, wherein the mitogen(s) is selected from the group consisting of EGE, bFGF, and betacellulin.

6. A method for obtaining human insulin secreting cells, comprising:

a) culturing hES cells in a first medium comprising Activin A;

b) culturing the cells from step a) in a second medium comprising a TGF-β

antagonist and one or more mitogens; and

c) culturing the cells from step b) in a third medium comprising nicotinamide;

thereby obtaining a population comprising human insulin-secreting cells.

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

antagonist is Noggin.

8. The method of claim 6, wherein the mitogen(s) is selected from the group consisting of epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), and betacellulin.

9. The method of claim 6, wherein the second medium comprises Noggin, EGF, and bFGF.

10. The method of claim 6, wherein the second medium comprises Noggin, betacellulin, and bFGF.