Stem cells and their use in transplantation
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.
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 ( βFGF-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 (βFGF-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.