Method of enriching for pancreatic endoderm cells

• US 9,506,034 B2
• Filed: 04/30/2015
• Issued: 11/29/2016
• 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:
  US14/700,608
• Publication Number:
  US 20150299662A1
• Time in Patent Office: 579 Days
• Field of Search: 435/325
• US Class Current: 1/1
• 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 endoderm cells comprising:

a) exposing an in vitro cell population comprising pancreatic endoderm cells to a ligand that binds CD142, CD55, CD57, or CD98; and

b) selecting cells in the in vitro cell population that express CD142 or CD57, or selecting cells in the in vitro cell population that do not express CD55 or CD98, thereby enriching for pancreatic endoderm cells.

2. The method of claim 1, wherein the pancreatic endoderm cells are derived from in vitro cultured pluripotent cells.

3. The method of claim 1, wherein the cells selected express CD142.

4. The method of claim 1, wherein the cells selected express CD57.

5. The method of claim 1, wherein the cells selected express CD55.

6. The method of claim 1, wherein the cells selected express CD98.

7. The method of claim 1, wherein the cells selected express CD142 but do not express CD55.

8. The method of claim 1, wherein the cells selected express CD142 but do not express CD98.

9. The method of claim 1, wherein greater than 50% of the pancreatic endoderm cells selected in step b) express CD142 and pancreatic and duodenal homeobox gene 1 (PDX1).

10. The method of claim 1, wherein greater than 50% of the pancreatic endoderm cells selected in step b) express CD57 and PDX1.

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

12. The method of claim 1, wherein the ligand comprises a detectable label.

13. The method of claim 1, wherein the ligand comprises a magnetic particle.

14. The method of claim 1, wherein the cells are selected by Fluorescence Activated Cell Sorting (FACS).