Stem cells and their use in transplantation

• US 7,438,902 B2
• Filed: 11/09/2004
• Issued: 10/21/2008
• Est. Priority Date: 12/06/1999
• Status: Expired due to Term

• Current Assignee: The General Hospital Corporation (Mass General Brigham, Inc.)
• Original Assignee: The General Hospital Corporation (Mass General Brigham, Inc.)
• Inventors: Faustman, Denise L., Thomas, Melissa K., Vallejo, Mario, Abraham, Elizabeth J., Habener, Joel E., Zulewski, Henryk
• Primary Examiner(s): Bunner; Bridget E
• Application Number:
  US10/984,645
• Publication Number:
  US 20050244386A1
• Time in Patent Office: 1,442 Days
• Field of Search: None
• US Class Current: 424/93.7
• 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: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• 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 5/0676 Pancreatic cells
• CPC Class Codes: C12N 5/0677 Three-dimensional culture, tissue culture or organ culture; Encapsulated cells
• 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 has been identified as a molecular marker for pancreatic stem cells, while cytokeratin-19 serves as a marker for a distinct class of islet ductal cells. Methods are described whereby nestin-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 method of treating a patient with diabetes mellitus, comprising the steps of:

(a) isolating a nestin-positive pancreatic stem cell from a pancreatic islet of a donor; and

(b) transferring the stem cell into the patient, wherein the stem cell differentiates into an insulin-producing cell,

wherein the patient does not serve as the donor for said stem cells of step (a),

thereby treating said patient with diabetes mellitus.

2. The method of claim 1, wherein the patient is a human and the donor for said stem cells of step (a) is a non-human animal.

3. The method of claim 1 or 2, wherein the patient is not treated with an immunosuppressive agent prior to step (b).

4. The method of claim 1 wherein, prior to the step of transferring, the stem cell is treated ex vivo with an agent selected from the group consisting of Epidermal Growth Factor (EGE), Basic Fibroblast Growth Factor-2 ( β

FGF-2 ), high glucose, Keratinocyte Growth Factor (KGF), Hepatocyte Growth Factor/Scatter Factor (HGF/SF), Glucagon-like Peptide-1(GLP-1 ), exendin-4, Islet/Duodenum Homeobox-1 (IDX-1 ), a nucleic acid molecule encoding IDX-1, betacellulin, activin A, Transforminci Growth Factor-β

(TGF-β

), and combinations thereof.

5. The method of claim 1, wherein the step of transferring is performed via endoscopic retrograde injection.

6. The method of claim 1 additionally comprising the step of:

(c) treating the patient with an immunosuppressive agent.

7. The method of claim 6 wherein said immunosuppressive agent prevents an immune response.

8. The method of claim 6 wherein said immunosuppressive agent delays the occurrence of an immune response.

9. The method of claim 6 wherein said immunosuppressive agent decreases the intensity of an immune response.

10. The method of claim 6, 7, 8 or 9 wherein the immune response is transplant rejection.

11. The method of claim 6, wherein the immunosuppressive agent is selected from the group consisting of FK-506, cyclosporin, and GAD65 antibodies.

12. A method of transplanting a stem cell into a mammal, comprising the steps of:

(a) isolating a nestin-positive pancreatic stem cell from a pancreatic islet of a donor; and

(b) transferring the stem cell into the mammal, wherein the stem cell differentiates into an insulin-producing cell.

13. The method of claim 12, wherein the mammal serves as the donor for said stem cells of step (a).

14. The method of claim 12, wherein the mammal does not serve as the donor for said stem cells of step (a).

15. The method of claim 12, wherein the mammal is a human and the donor for said stem cells of step (a) is a non-human mammal.

16. The method of claim 14 or 15, wherein the mammal is not treated with an immunosuppressive agent prior to step (b).

17. The method of claim 12 wherein, prior to the step of transferring, the stem cell is treated ex vivo with an agent selected from the group consisting of Epidermal Growth Factor (EGF), Basic Fibroblast Growth Factor-2 (β

FGF-2 ), high glucose, Keratinocvte Growth Factor (KGF), Hepatocyte Growth Factor/Scatter Factor (HG F/SF), Glucacion-like Peptide-1 (GLP-1), exendin-4, Islet/Duodenum Homeobox-1 (IDX-1 ), a nucleic acid molecule encoding IDX-1, betacellulin, activin A, Transforming Growth Factor-β

(TGF-β

), and combinations thereof.

18. The method of claim 12, wherein the step of transferring is performed via endoscopic retrograde injection.

19. The method of claim 12 additionally comprising the step of:

(c) treating the mammal with an immunosuppressive agent.

20. The method of claim 19 wherein said immunosuppressive agent prevents an immune response.

21. The method of claim 19 wherein said immunosuppressive agent delays the occurrence of an immune response.

22. The method of claim 19 wherein said immunosuppressive agent decreases the intensity of an immune response.

23. The method of claim 20, 21 or 22 wherein the immune response is transplant rejection.

24. The method of claim 19, wherein the immunosuppressive agent is selected from the group consisting of FK-506, cyclosporin, and GAD65 antibodies.

25. A method of treating a patient with diabetes mellitus, comprising the steps of:

(a) isolating a nestin-positive pancreatic stem cell from a pancreatic islet of a donor; and

(b) transferring the stem cell into the patient,

wherein the patient does not serve as the donor for said stem cells of step (a), thereby treating said patient with diabetes mellitus.

26. A method of transplanting a stem cell into a mammal, comprising the steps of:

(a) isolating a nestin-positive pancreatic stem cell from a pancreatic islet of a donor; and

(b) transferring the stem cell into the mammal.