Drug screening using islet cells and islet cell progenitors from human embryonic stem cells

• US 9,085,756 B2
• Filed: 04/19/2010
• Issued: 07/21/2015
• Est. Priority Date: 12/07/2001
• Status: Expired due to Fees

• Current Assignee: Asterias Biotherapeutics Incorporated (Lineage Cell Therapeutics, Inc.)
• Original Assignee: Asterias Biotherapeutics Incorporated (Lineage Cell Therapeutics, Inc.)
• Inventors: Fisk, Gregory J., Inokuma, Margaret S.
• Primary Examiner(s): Crouch, Deborah
• Application Number:
  US12/762,676
• Publication Number:
  US 20100196916A1
• Time in Patent Office: 1,919 Days
• US Class Current: 1/1
• 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. An in vitro method of screening a differentiation factor for its effect on differentiating endoderm cells that are the in vitro progeny of isolated primate pluripotent stem (pPS) cells into pancreatic islet cells, comprising

a) contacting a cell culture comprising gut endoderm cells expressing Sox17, HNF3β

and HNF4α

that are the in vitro progeny of isolated primate pluripotent stem (pPS) cells with a differentiation factor;

b) observing a change in the gut endoderm cells from step a) associated with the differentiation of an endoderm cell expressing Sox17, HNF3β and

HNF4α

into an islet cell; and

c) correlating the change in the gut endoderm cell with the factor.

2. The method of claim 1, wherein the gut endoderm cells are human cells.

3. The method of claim 1, wherein step b) comprises determining if the factor causes the gut endoderm cells to differentiate into a cell that expresses insulin.

4. The method of claim 1, wherein step b) comprises determining if the factor causes the endoderm pPS cells to differentiate into cells that expresses insulin c-peptide.

5. The method of claim 1, wherein the factor is a peptide.

6. The method of claim 1, wherein the factor is a growth factor.

7. The method of claim 1, wherein the factor is a morphogen.

8. An in vitro method of screening a factor chosen from a drug, a solvent, a peptide and a polynucleotide for its effect on primate pancreatic islet cells comprising

a) culturing primate pluripotent stem (pPS) cells in a first medium comprising Activin A to produce cells expressing Sox17, HNF3β

and HNF4α

;

b) maturing the cells from a) in a second medium comprising at least one differentiation factor selected from Activin A, cyclopamine, betacellulin, exendin-4, glucagon-like peptide 1 (GLP-1), hepatocyte growth factor (HGF), nicotinamide, insulin-like growth factor 1 (IGF-1), n-butyrate, retinoic acid, growth hormone, placental lactogen, vascular endothelial growth factor (VEGF), insulin-like growth factor II (IGF-II), 3-isobutyl-1-methylxanthine (IBMX), wortmannin, gastrin, cholecystokinin, nerve growth factor (NGF), epidermal growth factor (EGF), keratinocyte growth factor (KGF), platelet-derived growth factor (PDGF), regenerating gene (Reg), and islet neogenesis-associated protein (INGAP), to thereby to obtain primate pancreatic islet cells;

c) contacting the primate pancreatic islet cells with a test factor;

d) observing a change in the primate pancreatic islet cells after step c); and

e) correlating the change in the primate pancreatic islet cells with the factor.

9. The method of claim 8, wherein the primate pancreatic islet cells are human cells.

10. The method of claim 8, wherein the factor is a drug.

11. The method of claim 8, wherein the factor is a solvent.

12. The method of claim 8, wherein the factor is a peptide.

13. The method of claim 8, wherein the factor is a polynucleotide.

14. The method of claim 8, wherein the primate pancreatic islet cells express insulin.

15. The method of claim 8, wherein the primate pancreatic islet cells express insulin c-peptide.

16. The method of claim 8, wherein observing a change in the primate pancreatic islet cells comprises observing a morphological change in the primate pancreatic islet cells.

17. The method of claim 8, wherein observing a change in the primate pancreatic islet cells comprises observing a phenotypic change in the primate pancreatic islet.