METHODS FOR PURIFYING ENDODERM AND PANCREATIC ENDODERM CELLS DERIVED FROM HUMAN EMBRYONIC STEM CELLS

• US 20090263896A1
• Filed: 04/21/2008
• Published: 10/22/2009
• Est. Priority Date: 04/21/2008
• Status: Active Grant

• Current Assignee: Viacyte Incorporated (Vertex Pharmaceuticals Incorporated)
• Original Assignee: Viacyte Incorporated (Vertex Pharmaceuticals Incorporated)
• Inventors: Bang, Anne, Kelly, Olivia
• Granted Patent: US 8,338,170 B2
• Time in Patent Office: Days
• US Class Current: 435/366
• CPC Class Codes: C12N 2501/50 Cell markers; Cell surface determinants
• CPC Class Codes: C12N 2501/599 with CD designations not provided for elsewhere
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/0676 Pancreatic cells
• CPC Class Codes: C12N 5/0678 Stem cells; Progenitor cells; Precursor cells
• CPC Class Codes: G01N 33/56966 Animal cells
• CPC Class Codes: G01N 33/68 involving proteins, peptides or amino acids involving lipoproteins G01N33/92

Abstract

The present disclosure relates to compositions and methods comprising cell surface markers for hES-derived cells, in particular, endoderm lineage cells including pancreatic endoderm-type cells, derived from hES cells.

Claims

1. A purified cell population comprising human pancreatic endoderm cells.

2. The purified population of claim 1, wherein said human pancreatic endoderm cells comprise at least 50% of the human cells in said cell population.

3. The purified population of claim 1, wherein said human pancreatic endoderm cells comprise at least 75% of the human cells in said cell population.

4. The purified population of claim 1, wherein said human pancreatic endoderm cells express pancreatic and duodenal homeobox gene 1 (PDX1) and NKX6.1.

5. A method of purifying a pancreatic endoderm cell comprising:

a) exposing a population of cells derived from pluripotent stem cells comprising a pancreatic endoderm cell to a ligand which binds to a cell surface marker on the pancreatic endoderm cell, wherein the cell surface marker is selected from the group consisting of CD57 and CD142, and wherein the pancreatic endoderm cell expresses pancreatic and duodenal homeobox gene 1 (PDX1); and

b) separating the pancreatic endoderm cell from cells derived from pluripotent stem cells which do not bind to the ligand, thereby purifying said pancreatic endoderm cell.

6. The method of claim 5, wherein the ligand is an antibody or binding fragment thereof.

7. The method of claim 6, wherein said antibody is a monoclonal antibody.

8. The method of claim 6, wherein said antibody is a polyclonal antibody.

9. The method of claim 5, wherein the pancreatic endoderm cell expresses NKX6.1.

10. A method of purifying a pancreatic endoderm cell comprising:

a) exposing a population of cells derived from pluripotent stem cells comprising a pancreatic endoderm cell to a ligand which binds to a cell surface marker not expressed on said pancreatic endoderm cell, wherein said cell surface marker is selected from the group consisting of CD55 and CD98, and wherein the pancreatic endoderm cell expresses pancreatic and duodenal homeobox gene 1 (PDX1); and

b) separating the pancreatic endoderm cell from cells derived from pluripotent stem cells which bind to the ligand, thereby purifying said pancreatic endoderm cell.

11. The method of claim 10, wherein the ligand is an antibody or binding fragment thereof.

12. The method of claim 11, wherein said antibody is a monoclonal antibody.

13. The method of claim 11, wherein said antibody is a polyclonal antibody.

14. The method of claim 10, wherein the pancreatic endoderm cell expresses NKX6.1.

15. A method of purifying a pancreatic endocrine cell comprising:

a) exposing a population of cells derived from pluripotent stem cells comprising a pancreatic endocrine cell to a ligand which binds to a cell surface marker not expressed on said pancreatic endocrine cell, wherein the cell surface marker is selected from the group consisting of CD142 and CD340; and

b) separating the pancreatic endocrine cell from cells derived from pluripotent stem cells which bind to the ligand, thereby purifying said pancreatic endocrine cell.

16. The method of claim 15 wherein the pancreatic endocrine cell expresses at least chromogranin A (CHGA).

17. The method of claim 15, wherein the ligand is an antibody or binding fragment thereof.

18. The method of claim 17, wherein said antibody is a monoclonal antibody.

19. The method of claim 17, wherein said antibody is a polyclonal antibody.

20. A method of purifying a pancreatic endocrine cell comprising:

a) exposing a population of cells derived from pluripotent stem cells comprising a pancreatic endocrine cell to a ligand which binds to a cell surface marker expressed on the pancreatic endocrine cell, wherein the cell surface marker is selected from the group consisting of CD57, CD200, and CD318; and

b) separating the pancreatic endocrine cell from cells derived from pluripotent stem cells which do not bind to the ligand, thereby purifying said pancreatic endocrine cell.

21. The method of claim 20, wherein the pancreatic endocrine cell expresses at least chromogranin A (CHGA).

22. The method of claim 20, wherein the ligand is an antibody or binding fragment thereof.

23. The method of claim 22, wherein said antibody is a monoclonal antibody.

24. The method of claim 22, wherein said antibody is a polyclonal antibody.