Compositions and methods for promoting the generation of PDX1+ pancreatic cells

• US 8,728,812 B2
• Filed: 04/22/2009
• Issued: 05/20/2014
• 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: Chen, Shuibing, Melton, Douglas A., Borowiak, Malgorzata, Lamenzo, Julia, Schreiber, Stuart L., Peng, Lee F., Davidow, Lance, Lam, Kelvin, Rubin, Lee L.
• Primary Examiner(s): Singh, Anoop
• Assistant Examiner(s): Montanari, David A
• Application Number:
  US12/989,284
• Publication Number:
  US 20110070645A1
• Time in Patent Office: 1,854 Days
• Field of Search: 435/325, 435/366, 435/377
• 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 Pdx1 expressing cells comprising:

a) contacting a population of human embryonic stem cells with Activin A to differentiate them into definitive endoderm cells, and

b) exposing the definitive endoderm cells of step a) to an effective amount of at least one compound listed in table 1 to increase the number of Pdx1 expressing cells in the differentiated cell population.

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

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

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

5. A kit comprising:

human embryonic stem cells;

Activin A;

at least one compound listed in table 1; and

instructions for differentiating the stem cells to definitive endoderm using the Activin A and for using the at least one compound to increase the number of Pdx1 expressing cells in a differentiated cell population.

6. The kit of claim 5, in which the compound is compound 1 in table 1.

7. The kit of claim 6, further comprising at least one additional agent.

8. The kit of claim 7, in which the at least one additional agent is FGF10.

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

a) contacting human embryonic stem cells with Activin A to produce definitive endoderm cells; and

b) contacting the definitive endoderm cells of with an effective amount of at least one compound listed in table 1 to increase the number and percentage of Pdx 1+pancreatic precursors.

10. The method of claim 9, in which the at least one compound is compound 1 in table 1.

11. The method of claim 10, further comprising differentiating the Pdx1+pancreatic precursors into mature pancreatic cells.