Generation of anterior foregut endoderm from pluripotent cells

• US 9,234,170 B2
• Filed: 04/25/2011
• Issued: 01/12/2016
• Est. Priority Date: 04/25/2010
• Status: Active Grant

• Current Assignee: Mount Sinai School of Medicine (Sinai Health System)
• Original Assignee: Mount Sinai School of Medicine (Sinai Health System)
• Inventors: Snoeck, Hans-Willem, Green, Michael
• Primary Examiner(s): Kim, Taeyoon
• Assistant Examiner(s): Patury, Srikanth
• Application Number:
  US13/643,032
• Publication Number:
  US 20130217005A1
• Time in Patent Office: 1,723 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• 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 2501/119 Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• 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 2501/385 of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR]
• CPC Class Codes: C12N 2501/40 Regulators of development
• CPC Class Codes: C12N 2501/41 Hedgehog proteins; Cyclopamine (inhibitor)
• CPC Class Codes: C12N 2501/415 Wnt; Frizzeled
• 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 2506/45 from artificially induced pluripotent stem cells
• CPC Class Codes: C12N 5/0603 Embryonic cells production of embryos, nuclear transfer A01K67/027; Embryoid bodies
• CPC Class Codes: C12N 5/0607 Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0617 Thyroid and parathyroid glands
• CPC Class Codes: C12N 5/0688 Cells from the lungs or the respiratory tract

Abstract

The invention is directed to in vitro methods of inducing differentiation of anterior foregut endoderm and the enriched populations of anterior foregut endoderm produced by such methods. Such enriched populations are useful for studies of the molecular events that occur during differentiation and for generating cells for cell replacement therapy.

Claims

1. A method of deriving an anterior foregut endoderm cell, the method comprising culturing a definitive endoderm cell derived from a pluripotent cell, in the presence of an inhibitor of bone morphogenetic protein (BMP) and in the presence of an inhibitor of transforming growth factor beta (TGF-beta) signaling.

2. The method of claim 1, wherein the definitive endoderm cell is cultured in the absence of Activin A.

3. The method of claim 1, wherein the inhibitor of BMP is Noggin, Chordin, or follistatin.

4. The method of claim 3, wherein the inhibitor of BMP is Noggin.

5. The method of claim 1, wherein the inhibitor of TGF-beta signaling is SB-431542.

6. The method of claim 2, wherein the BMP inhibitor is Noggin and the inhibitor of TGF-beta signaling is SB-431542.

7. The method of claim 1, wherein the definitive endoderm cell is an embryoid body cell.

8. The method of claim 1, wherein the anterior foregut endoderm cell is a human cell.

9. The method of claim 1, wherein the pluripotent cell is an embryonic stem cell.

10. The method of claim 1, wherein the pluripotent cell is an induced pluripotent stem cell.

11. The method of claim 1, wherein the pluripotent cell is a human cell.

12. The method of claim 1, wherein the anterior foregut endoderm cell expresses Foxa2 and comprises an elevated level of Sox2 expression and a reduced level of Cdx2 expression, compared to the expression of Sox2 and Cdx2 in the definitive endoderm cell.