Method of enriching for pancreatic endocrine cells that express chromogranin A

• US 10,266,808 B2
• Filed: 10/28/2016
• Issued: 04/23/2019
• 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
• Primary Examiner(s): Wilson, Michael C
• Application Number:
  US15/338,149
• Publication Number:
  US 20170044498A1
• Time in Patent Office: 907 Days
• Field of Search: 435325
• 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 that express chromogranin A (CHGA), the method comprising:

a) exposing an in vitro population of pancreatic cells to a ligand that binds CD200, CD318, CD57, CD142, or CD340; and

b) either i) selecting cells that bind to the ligand that binds CD200, CD318, or CD57;

or

ii) selecting cells that do not bind to the ligand that binds CD142 or CD340,

thereby enriching for pancreatic endocrine cells that express CHGA.

2. The method of claim 1, wherein said ligand comprises an antibody or binding fragment thereof that specifically binds the cell-surface marker.

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

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

5. The method of claim 1, wherein the selecting is performed by Fluorescence Activated Cell Sorting (FACS).

6. The method of claim 1, comprising selecting cells that bind to the ligand that binds CD318, thereby enriching for pancreatic endocrine cells that express CHGA and insulin.

7. A method of enriching for pancreatic endocrine cells that express chromogranin A (CHGA), the method comprising:

a) exposing an in vitro population of pancreatic cells to a ligand that binds CD200, CD318, or CD57; and

b) selecting cells that bind to the ligand that binds CD200, CD318, or CD57,

thereby enriching for pancreatic endocrine cells that express CHGA.

8. A method of enriching for pancreatic endocrine cells that express chromogranin A (CHGA), the method comprising:

a) exposing an in vitro population of pancreatic cells to a ligand that binds CD142, or CD340; and

b) selecting cells that do not bind to the ligand that binds CD142 or CD340,

thereby enriching for pancreatic endocrine cells that express CHGA.