Methods for differentiating and purifying pancreatic endocrine cells

Patent metadata

Field	Value
Publication number	US-11679133-B2
Application number	US-201615780153-A
Country	US
Kind code	B2
Filing date	Nov 28, 2016
Priority date	Nov 30, 2015
Publication date	Jun 20, 2023
Grant date	Jun 20, 2023

Abstract

The present invention relates to compositions and methods comprising cell surface markers for pluripotent-derived cells, in particular, pancreatic endoderm-type cells, derived from pluripotent stem cells.

First claim

What is claimed is:

1. A method of enriching for pancreatic endocrine cells expressing NKX6.1 and insulin comprising:
a) exposing an in vitro cell population comprising pancreatic endocrine cells expressing NKX6.1 and insulin having been ex-vivo differentiated from pluripotent stem cells to a ligand that binds CD49a;
b) exposing the in vitro cell population comprising pancreatic endocrine cells expressing NKX6.1 and insulin having been ex-vivo differentiated from pluripotent stem cells to a ligand that binds CD26;
c) selecting cells which bind said ligand that binds CD49a and do not bind said ligand that binds CD26; and subsequently
d) culturing said cells which bind said ligand that binds CD49a and do not bind said ligand that binds CD26, thereby enriching for pancreatic endocrine cells expressing NKX6.1 and insulin.

2. The method of claim 1 , wherein the ligand that binds CD49a is an antibody or binding fragment thereof.

3. The method of claim 2 , wherein said antibody is a monoclonal antibody.

4. The method of claim 1 , wherein the ligand that binds CD49a is associated with a magnetic particle.

5. The method of claim 1 , wherein the selecting is by fluorescence-based cell sorting or magnetic-field based cell sorting.

6. The method of claim 1 , further comprising ex vivo differentiating pancreatic endocrine cells from human pluripotent stem cells prior to step (a).

Assignees

• Kadimastem Ltd
• Yeda Res & Dev

Inventors

• Molakandov Kfir
• Lavon Neta
• Beck Avital
• Revel Michel
• Elhanani Ofer
• Soen Yoav
• Walker Michael
• Hasson Arik

Classifications

• C12N2500/34

  Sugars · CPC title

• A61K35/39 Primary

  Pancreas; Islets of Langerhans (Langerhans cells of epidermis A61K35/36) · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• C12N5/0676

  Pancreatic cells · CPC title

• C12N2500/38

  Vitamins · CPC title