Populations of pancreatic progenitor cells and methods of isolating and using same

• US 8,927,274 B2
• Filed: 04/12/2011
• Issued: 01/06/2015
• Est. Priority Date: 04/12/2010
• Status: Expired due to Fees

• Current Assignee: TECHNION RESEARCH AND DEVELOPMENT FOUNDATION, LTD. (Technion-Israel Institute Of Technology)
• Original Assignee: TECHNION RESEARCH AND DEVELOPMENT FOUNDATION, LTD. (Technion-Israel Institute Of Technology)
• Inventors: Itskovitz-Eldor, Joseph, Fishman, Bettina, Segev, Hanna
• Primary Examiner(s): Tate, Chris R
• Assistant Examiner(s): Kosar, Aaron J
• Application Number:
  US13/640,532
• Publication Number:
  US 20130034526A1
• Time in Patent Office: 1,365 Days
• Field of Search: None
• US Class Current: 435/366
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2506/45 from artificially induced pluripotent stem cells
• CPC Class Codes: C12N 5/0678 Stem cells; Progenitor cells; Precursor cells

Abstract

A method of generating pancreatic progenitor cells is disclosed. The method comprises:

• • (a) differentiating stem cells under conditions such that at least a portion of the cells express glucose transporter 2 (GLUT2) so as to generate GLUT2-expressing cells; and
  • (b) enriching for the GLUT2-expressing cells so as to generate a population of GLUT2 enriched cells, wherein at least 80% of the population of GLUT2 enriched cells express GLUT2, thereby generating pancreatic progenitor cells.

Isolated populations of cells generated according to the method, pharmaceutical compositions comprising same and uses thereof are also disclosed.

Claims

1. A method of generating GLUT2-expressing cells the method comprising:

(a) differentiating human stem cells under conditions such that at least a portion of said cells express glucose transporter 2 (GLUT2) on a membrane of said cells so as to generate GLUT2-expressing cells; and

(b) isolating from said cells resultant of step (a) GLUT2-expressing cells using an anti GLUT2 antibody which binds to said GLUT2 expressed on said membrane so as to generate a population of GLUT2 enriched cells, wherein at least 80% of said population of GLUT2 enriched cells express GLUT2, thereby generating GLUT2-expressing cells.

2. The method of claim 1, wherein said isolating is effected by a fluorescence-activated cell sorter (FACS).

3. The method of claim 1, wherein said human stem cells comprise human embryonic stem cells.

4. The method of claim 1, wherein said stem cells comprise induced pluripotent stem cells.

5. The method of claim 1, wherein said human stem cells comprise human pluripotent stem cells.

6. The method of claim 1, wherein said differentiating said human stem cells is effected by generating embryoid bodies.

7. The method of claim 6, wherein said embryoid bodies are 14 day old.

8. The method of claim 6, wherein said embryoid bodies 14-25 day old.

9. The method of claim 6, wherein said embryoid bodies are 14-19 day old.

10. The method of claim 1, wherein said conditions comprise a glucose concentration of less than 5 mM.

11. The method of claim 1, wherein said isolating is effected by a magnetic-activated cell sorting (MACS).

12. The method of claim 1, further comprising:

(c) differentiating said population of GLUT2 enriched cells into insulin-producing cells.