COMPOSITIONS AND METHODS FOR PROMOTING THE GENERATION OF PDX1+ PANCREATIC CELLS

• US 20110070645A1
• Filed: 04/22/2009
• Published: 03/24/2011
• Est. Priority Date: 04/22/2008
• Status: Active Grant

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation), The General Hospital Corporation (Mass General Brigham, Inc.)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation), The General Hospital Corporation (Mass General Brigham, Inc.)
• Inventors: Borowiak, Malgorzata, Davidow, Lance, Lam, Kelvin, Chen, Shuibing, Peng, Lee F., Lamenzo, Julia, Rubin, Lee L., Melton, Douglas A., Schreiber, Stuart L.
• Granted Patent: US 8,728,812 B2
• Time in Patent Office: Days
• US Class Current: 435/377
• 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 2501/16 Activin; Inhibin; Mullerian inhibiting substance
• 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 2501/415 Wnt; Frizzeled
• CPC Class Codes: C12N 2501/42 Notch; Delta; Jagged; Serrate
• 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 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

Abstract

Certain embodiments disclosed herein are directed to a method of producing pancreatic cells or pancreatic cell precursors by exposing human embryonic stem cells to an effective amount of at least one compound listed in Table I to differentiate the human embryonic stem cells into the pancreatic cells or the pancreatic cell precursors. Kits and pancreatic cell lines produced using the methods are also described.

Claims

1. A method of producing pancreatic cells or pancreatic cell precursors comprising exposing human embryonic stem cells to an effective amount of at least one compound listed in Table I to differentiate the human embryonic stem cells into the pancreatic cells or the pancreatic cell precursors.

2. The method of claim 1, in which the compound is Compound I in Table I

3. The method of claim 2, further comprising exposing the human embryonic stem cell to at least one additional agent.

4. The method of claim 3, in which the at least one additional agent is FGF10.

5. The method of claim 1, in which the human embryonic stem cells are first differentiated to endoderm and then exposed to the at least one compound listed in Table Ito provide the pancreatic cells or the pancreatic cell precursors.

34. The method of claim 1, wherein the pancreatic precursor cells are Pdx1+.

6. A pancreatic cell or pancreatic cell precursor produced by exposing a human embryonic stem cell to at least one compound listed in Table I.

7. The pancreatic cell or pancreatic cell precursor of claim 6, in which the compound is Compound I in Table I.

8. The pancreatic cell or pancreatic cell precursor of claim 7, in which the pancreatic cell or pancreatic cell precursor is produced by exposing the stem cell to the at least one compound in combination with at least one additional agent.

9. The pancreatic cell or pancreatic cell precursor of claim 8, in which the at least one additional agent is FGF10.

10. The pancreatic cell of pancreatic cell precursor of claim 6, in which the human embryonic stem cells are first differentiated to endoderm prior to exposure to the at least one compound listed in Table I.

35. The method of claim 6, wherein the pancreatic precursor cells are Pdx1+.

11. A kit comprising:

human embryonic stem cells;

at least one compound listed in Table I; and

instructions for using the stem cells and the at least one compound to produce a pancreatic cell or a pancreatic cell precursor.

12. The kit of claim 11, in which the compound is Compound I in Table I.

13. The kit of claim 12, further comprising at least one additional agent.

14. The kit of claim 13, in which the at least one additional agent is FGF10.

15. The kit of claim 11, in which the human embryonic stem cells are first differentiated to endoderm prior to packaging in the kit.

16-30. -30. (canceled)

31. A method of producing Pdx1+ pancreatic precursors comprising:

differentiating human embryonic stem cells to provide endoderm; and

culturing the endoderm in the presence of an effective amount of at least one compound listed in Table Ito produce the Pdx1+ pancreatic precursors.

32. The method of claim 31, in which the at least one compound is Compound I in Table I.

33. The method of claim 32, further comprising differentiating the Pdx1+ pancreatic precursors into mature pancreatic cells.