SC-BETA CELLS AND COMPOSITIONS AND METHODS FOR GENERATING THE SAME

• US 20150240212A1
• Filed: 04/10/2015
• Published: 08/27/2015
• Est. Priority Date: 06/11/2013
• Status: Abandoned Application

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Peterson, Quinn P., Pagliuca, Felicia J., Melton, Douglas A., Millman, Jeffrey R., Segel, Michael Saris, Gurtler, Mads
• Application Number:
  US14/684,129
• Publication Number:
  US 20150240212A1
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 5/48 of the pancreatic hormones
• CPC Class Codes: A61P 5/50 for increasing or potentiating the activity of insulin
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• 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 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/16 Activin; Inhibin; Mullerian inhibiting substance
• CPC Class Codes: C12N 2501/375 Thyroid stimulating hormone [TSH]
• 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 2501/395 Thyroid hormones
• CPC Class Codes: C12N 2501/40 Regulators of development
• CPC Class Codes: C12N 2501/41 Hedgehog proteins; Cyclopamine (inhibitor)
• CPC Class Codes: C12N 2501/727 Kinases (EC 2.7.)
• CPC Class Codes: C12N 2501/998 Proteins not provided for elsewhere
• 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 2506/03 from non-embryonic pluripotent stem cells
• CPC Class Codes: C12N 2506/22 from pancreatic cells
• CPC Class Codes: C12N 2506/45 from artificially induced pluripotent stem cells
• 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 5/0676 : Pancreatic cells
• CPC Class Codes: C12N 5/0678 : Stem cells; Progenitor cells; Precursor cells
• CPC Class Codes: C12N 5/0696 : Artificially induced pluripotent stem cells, e.g. iPS

Abstract

Disclosed herein are methods, compositions, kits, and agents useful for inducing β cell maturation, and isolated populations of SC-β cells for use in various applications, such as cell therapy.

Claims

1. A method of differentiating progenitor cells into pancreatic β

cells in vitro, the method comprising culturing a cluster of the progenitor cells in suspension in a culture medium comprising the following compounds;

a thyroid hormone signaling pathway activator and a transforming growth factor β

(TGF-β

) signaling pathway inhibitor, thereby differentiating the progenitor cells into pancreatic β

cells, wherein the pancreatic β

cells exhibit an in vitro glucose-stimulated insulin secretion response to at least a first glucose challenge and a second glucose challenge when the first glucose challenge and the second glucose challenge are applied sequentially.

2. The method of claim 1, wherein the progenitor cells comprise an insulin-positive endocrine cell.

3. The method of claim 1, wherein the progenitor cells comprise a PDX1-positive progenitor cell.

4. The method of claim 1, wherein the thyroid hormone signaling pathway activator is triiodothyronine.

5. The method of claim 4, wherein the concentration of the triiodothyronine is at least 0.1 μ

M.

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

signaling pathway inhibitor is Alk5 inhibitor II.

7. The method of claim 6, wherein the concentration of the Alk5 inhibitor II is at least 100 nM.

8. The method of claim 1, wherein the culturing comprises changing the culture medium at least every other day.

9. The method of claim 1, wherein the culturing lasts for at least seven days.

10. The method of claim 1, wherein the culture medium is a serum-free culture medium.

11. The method of claim 1, wherein the progenitor cells are cultured in the presence of a low-adherent substrate.

12. The method of claim 1, wherein the progenitor cells are human progenitor cells.

13. The method of claim 1, wherein the progenitor cells are generated from stem cells.

14. The method of claim 13, wherein the stem cells are embryonic stem cells.

15. The method of claim 13, wherein the stem cells are induced pluripotent stem cells.

16. The method of claim 1, wherein the non-native pancreatic β

cells exhibit an in vitro glucose-stimulated insulin secretion response to a first glucose challenge, a second glucose challenge, and a third glucose challenge, when the first glucose challenge, the second glucose challenge and the third glucose challenge are applied sequentially.