Stem cells of the islets of langerhans and their use in treating diabetes mellitus

• US 8,110,399 B2
• Filed: 05/22/2006
• Issued: 02/07/2012
• Est. Priority Date: 10/06/2000
• Status: Expired due to Fees

• Current Assignee: General Hospital Corporation, The General Hospital Corporation (Mass General Brigham, Inc.)
• Original Assignee: The General Hospital Corporation (Mass General Brigham, Inc.)
• Inventors: Habener, Joel F., Zulewski, Henryk, Thomas, Melissa K., Abraham, Elizabeth J., Vallejo, Mario, Nolan, Anna Louise, Leech, Colin A., Lechner, Andreas
• Primary Examiner(s): Kelly, Robert M
• Application Number:
  US11/438,790
• Publication Number:
  US 20070128176A1
• Time in Patent Office: 2,087 Days
• Field of Search: None
• US Class Current: 435/366
• CPC Class Codes: A01K 67/0271 Chimeric vertebrates, e.g. comprising exogenous cells
• CPC Class Codes: A61K 2035/122 for inducing tolerance or supression of immune responses
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C12N 2500/34 Sugars
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/335 Glucagon; Glucagon-like peptide [GLP]; Exendin
• CPC Class Codes: C12N 2510/02 Cells for production
• CPC Class Codes: C12N 5/0678 Stem cells; Progenitor cells; Precursor cells

Abstract

Methods and compositions are described for the treatment of type I insulin-dependent diabetes mellitus and other conditions using newly identified stem cells that are capable of differentiation into a variety of pancreatic islet cells, including insulin-producing beta cells, as well as hepatocytes. Nestin and ABCG2 have been identified as molecular markers for pancreatic stem cells, while cytokeratin-19 serves as a marker for a distinct class of islet ductal cells. Methods are described whereby nestin and/or ABCG2-positive stem cells can be isolated from pancreatic islets and cultured to obtain further stem cells or pseudo-islet like structures. Methods for ex vivo differentiation of the pancreatic stem cells are disclosed. Methods are described whereby pancreatic stem cells can be isolated, expanded, and transplanted into a patient in need thereof, either allogeneically, isogeneically or xenogenically, to provide replacement for lost or damaged insulin-secreting cells or other cells.

Claims

1. A population of nestin-positive human pancreatic stem cells wherein said nestin-positive stem cells are also positive for at least one of GLP-1 R, ABCG2, integrin subunit α

6 and integrin subunit β

1;

wherein said nestin-positive stem cells of said population are not neural stem cells,

wherein said population is at least 30% pure; and

wherein said nestin-positive stem cells grow in RPMI-1640 .

2. The population of nestin-positive stem cells of claim 1, wherein said nestin-positive stem cells are also positive for at least one of the markers selected from the group consisting of Oct3/4, latrophilin (type 2), Hes-1, C-kit, MDR-1, SUR-1, or Kir 6.2.

3. The population of nestin-positive stem cells of claim 1, wherein said nestin-positive cells do not express at least one of the markers selected from the group consisting of CD34, CD45, CD133, MHC class I and MHC class II.

4. The population of nestin-positive stem cells of claim 1, wherein said nestin-positive stem cells differentiate to form insulin-producing beta cells.

5. The population of nestin-positive stem cells of claim 1, wherein said nestin-positive stem cells differentiate to form glucagon-producing alpha cells.

6. The population of nestin-positive stem cells of claim 1, wherein said nestin-positive stem cells differentiate to form pseudo-islet like aggregates.

7. A pharmaceutical composition comprising nestin-positive stem cell of the population of nestin-positive stem cells of claim 1 admixed with a physiologically compatible carrier.

8. A population of nestin-positive human pancreatic stem cells wherein said nestin-positive stem cells are also positive for at least one of GLP-1 R, ABCG2, integrin subunit α

6 and integrin subunit β

1;

wherein said nestin-positive stem cells are not neural stem cells;

wherein said stem cell population is at least 40% pure; and

wherein said nestin-positive stem cells grow in RPMI-1640.

9. A population of nestin-positive human pancreatic stem cells that wherein said nestin-positive stem cells are also positive for at least one of GLP-1 R, ABCG2, integrin subunit α

6 and integrin subunit β

1;

wherein said nestin-positive stem cells are not neural stem cells;

wherein said population is at least 50% pure; and

wherein said nestin-positive stem cells grow in RPMI-1640.

10. A population of nestin-positive human pancreatic stem cells wherein said nestin-positive stem cells are also positive for at least one of GLP-1 R, ABCG2;

wherein said nestin-positive stem cells are not neural stem cells;

wherein said population is at least 60% pure; and

wherein said nestin-positive stem cells grow in RPMI-1640.

11. A population of nestin-positive human pancreatic stem cells wherein said nestin-positive stem cells are also positive for at least one of GLP-1 R, ABCG2, integrin subunit α

6 and integrin subunit β

1;

wherein said nestin-positive stem cells are not neural stem cells;

wherein said population is at least 70% pure; and

wherein said nestin-positive stem cells grow in RPMI-1640.

12. A population of nestin-positive human pancreatic stem cells that wherein said nestin-positive stem cells are also positive for at least one of GLP-1 R, ABCG2, integrin subunit α

6 and integrin subunit β

1;

wherein said nestin-positive stem cells are not neural stem cells;

wherein said population is at least 80% pure; and

wherein said nestin-positive stem cells grow in RPMI-1640.

13. A population of nestin-positive human pancreatic stem cells wherein said nestin-positive stem cells are also positive for at least one of GLP-1 R or ABCG2, integrin subunit α

6 and integrin subunit β

1;

wherein said nestin-positive stem cells are not neural stem cells;

wherein said population is at least 85% pure; and

wherein said nestin-positive stem cells grow in RPMI-1640.

14. A population of nestin-positive human pancreatic stem cells wherein said nestin-positive stem cells are also positive for at least one of GLP-1 R or ABCG2, integrin subunit α

6 and integrin subunit β

1;

wherein said nestin-positive stem cells are not neural stem cells;

wherein said population is at least 90% pure; and

wherein said nestin-positive stem cells grow in RPMI-1640.

15. A population of nestin-positive human pancreatic stem cells wherein said nestin-positive stem cells are also positive for at least one of GLP-1 R, ABCG2, integrin subunit α

6 and integrin subunit β

1;

wherein said nestin-positive stem cells are not neural stem cells;

wherein said population is at least 95% pure; and

wherein said nestin-positive stem cells grow in RPMI-1640.

16. A population of nestin-positive human pancreatic stem cells wherein said nestin-positive stem cells are also positive for at least one of GLP-1 R, ABCG2, integrin subunit α

6 and integrin subunit β

1;

wherein said nestin-positive stem cells are not neural stem cells;

wherein said population is at least 98% pure; and

wherein said nestin-positive stem cells grow in RPMI-1640.

17. A population of nestin-positive human pancreatic stem cells wherein said nestin-positive stem cells are also positive for at least one of GLP-1R, ABCG2, integrin subunit α

6 and integrin subunit β

1;

wherein said nestin-positive stem cells are not neural stem cells;

wherein said population is at least 99% pure; and

wherein said nestin-positive stem cells grow in RPMI-1640.