Production of insulin producing cells

• US 9,624,472 B2
• Filed: 05/18/2015
• Issued: 04/18/2017
• Est. Priority Date: 06/17/2010
• Status: Active Grant

• Current Assignee: Regents of The University of Minnesota (University of Minnesota)
• Original Assignee: Regents of The University of Minnesota (University of Minnesota)
• Inventors: Firpo, Meri, Geng, Zhaohui
• Primary Examiner(s): Shen, Bin
• Application Number:
  US14/715,098
• Publication Number:
  US 20150322407A1
• Time in Patent Office: 701 Days
• 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: C12N 2501/11 Epidermal growth factor [EGF]
• 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 2501/335 Glucagon; Glucagon-like peptide [GLP]; Exendin
• CPC Class Codes: C12N 2501/415 Wnt; Frizzeled
• CPC Class Codes: C12N 2501/91 Heparin
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2506/45 from artificially induced pluripotent stem cells
• CPC Class Codes: C12N 5/0676 Pancreatic cells
• CPC Class Codes: C12N 5/0678 Stem cells; Progenitor cells; Precursor cells

Abstract

The invention provides methods for differentiating stem cells along the pancreatic lineage as well as large scale culture methods. The present invention further provides pancreatic progenitor cells derived from stem cells to provide pancreatic cells to a subject.

Claims

1. A method to provide pancreatic cells to a subject in need thereof comprising:

a) contacting the stem cells with at least one member of the TGFβ

family of cytokines and at least one member of the Wnt family of proteins, wherein the stem cells are induced pluripotent stem (iPS) cells,

b) contacting the cells obtained from step a) with at least one member of the TGFβ

family of cytokines, at least one member of the Wnt family of proteins, and an agent that inhibits sonic hedgehog activity; and

c) contacting the cells obtained from step b) with a member of the Epidermal growth factor (EGF) family of proteins; and

administering the cells so as to provide pancreatic cells in the subject.

2. The method of claim 1, wherein the at least one member of the TGFβ

family of cytokines is activinA or nodal.

3. The method of claim 1, wherein the at east one member of the Wnt family is Wnt3 or Wnt3A.

4. The method of claim 1, wherein the at least one member of the EGF family is EGF.

5. The method of claim 1 further comprising contacting the cells obtained from step c) with at least one member of the TGFβ

family of cytokines, at least one member of the Wnt family of proteins, exendin4 or a combination thereof to yield cells expressing insulin prior to administration to the subject.

6. The method of claim 5, wherein the at least one member of the TGFβ

family of cytokines is GDF-11.

7. The method of claim 5, wherein the at least one member of the Wnt family or proteins is betacellulin.

8. The method of claim 1, wherein the subject is a mammal.

9. The method of claim 8, wherein the mammal is a human.

10. The method of claim 1, wherein the subject has a pancreatic disorder or injury.

11. The method of claim 10, wherein the disorder comprises diabetes, obesity, pancreatic atresia, pancreas inflammation, alpha1-antitrypsin deficiency, hereditary pancreatitis, pancreatic cancer, pancreatic enzyme deficiency or hyperinsulinism.

12. The method of claim 11, wherein the diabetes is Type I or Type II diabetes.

13. The method of claim 10, wherein the injury is a result of physical trauma, chemical, radiation, aging, disease or combination thereof.

14. A method to provide pancreatic cells to a subject in need thereof comprising:

a) contacting stem cells with Activin A and Wnt 3a, wherein the stem cells are induced pluripotent stem (iPS) cells;

b) contacting the cells obtained from step a) with Activin-A, Wnt3a, and an agent that inhibits sonic hedgehog activity;

c) contacting the cells obtained from step b) with EGF; and

administering the cells so as to provide pancreatic cells in the subject.

15. A method to provide insulin expressing cells to a subject in need thereof comprising:

a) contacting stem cells with Activin A and Wnt3a, wherein the stem cells are induced pluripotent stem (iPS) cells;

b) contacting the cells obtained from step a) with Activin-A, Wnt3a, and an agent that inhibits sonic hedgehog activity;

c) contacting the cells obtained from step b) with EGF;

d) contacting the cells obtained from step c) with GDF-11, betacellulin, exendin4 or a combination thereof so as to yield cells expressing insulin or having increased expression of insulin; and

e) administering the cells expressing insulin or having increased expression of insulin to the subject.