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

• US 20130122520A1
• Filed: 12/21/2012
• Published: 05/16/2013
• Est. Priority Date: 04/21/2008
• Status: Active Grant

• Current Assignee: Viacyte Incorporated (Vertex Pharmaceuticals Incorporated)
• Original Assignee: Viacyte Incorporated (Vertex Pharmaceuticals Incorporated)
• Inventors: Kelly, Olivia, Bang, Anne
• Granted Patent: US 9,045,736 B2
• Time in Patent Office: Days
• US Class Current: 435/7.21
• 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 method of enriching for pancreatic endocrine cells comprising:

a) exposing an in vitro cell population comprising pancreatic endocrine cells to a ligand that binds a cell-surface marker selected from the group consisting of CD200, CD318, CD142, CD57 and CD340; and

b) selecting cells that bind to said ligand that binds the marker selected from the group consisting of CD200, CD318 and CD57 or selecting cells that do not bind to said ligand that binds the marker selected from the group consisting of CD142 and CD340, thereby enriching for pancreatic endocrine cells.

2. The method of claim 1, wherein said enriched pancreatic endocrine cells express chromogranin A.

3. The method of claim 1, wherein said enriched pancreatic endocrine cells express insulin.

4. The method of claim 1, wherein the ligand comprises an antibody or binding fragment thereof.

5. The method of claim 1, wherein said ligand is associated with a detectable label.

6. The method of claim 1, wherein said ligand is associated with a magnetic particle.

7. The method of claim 1, wherein said cells are separated by Fluorescence Activated Cell Sorting (FACS).

8. The method of claim 1, wherein said ligand binds CD318.

9. The method of claim 8, wherein the ligand comprises an antibody or binding fragment thereof.

10. The method of claim 1, wherein said ligand binds CD200.

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

12. The method of claim 1, wherein said ligand binds CD57.

13. The method of claim 12, wherein the ligand comprises an antibody or binding fragment thereof.

14. The method of claim 1, wherein said ligand binds CD340.

15. The method of claim 14, wherein the ligand comprises an antibody or binding fragment thereof.

16. The method of claim 1, wherein said ligand binds CD142.

17. The method of claim 16, wherein the ligand comprises an antibody or binding fragment thereof.

18. A method of enriching for cells expressing chromogranin A comprising:

a) exposing an in vitro cell population comprising chromogranin A positive cells to a ligand that binds a cell-surface marker selected from the group consisting of CD200, CD318, CD142, CD57 and CD340; and

b) selecting cells that bind to said ligand that binds the marker selected from the group consisting of CD200, CD318 and CD57 or selecting cells that do not bind to said ligand that binds the marker selected from the group consisting of CD142 and CD340, thereby enriching for cells expressing chromogranin A.

19. A cell culture comprising endocrine cells that express cell surface markers selected from the group consisting of CD200, CD318 and CD57 and that do not express cell surface markers selected from the group consisting of CD142 and CD340.