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;
(b) culturing the stem cell ex vivo to produce a progenitor cell; and
(c) transferring the progenitor cell into the patient, wherein the progenitor cell differentiates into an insulin-producing beta 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. 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;
(b) expanding the stem cell ex vivo to produce a progenitor cell; and
(c) transferring the progenitor cell into the patient, wherein the progenitor cell differentiates into an insulin-producing beta cell,
wherein the patient does not serve as the donor for said stem cells of step (a);
thereby treating said patient with diabetes mellitus.
3. The method of claim 1 or 2 , wherein the patient is a human and the donor for said stem cells of step (a) is a non-human mammal.
4. The method of claim 1 or 2 , wherein the patient is not treated with an immunosuppressive agent prior to step (b).
5. The method of claim 2 , wherein the step of expanding is performed in the presence of an agent selected from the group consisting of EGF, bFGF-2, high glucose, KGF, HGF/SF, GLP-1, exendin-4, IDX-1, a nucleic acid molecule encoding IDX-1, betacellulin, activin A, TGF-β, and combinations thereof.
6. The method of claim 1 or 2 , wherein the step of transferring is performed via endoscopic retrograde injection.
7. The method of claim 1 additionally comprising the step of:
(d) treating the patient with an immunosuppressive agent.
8. The method of claim 2 additionally comprising the step of:
(d) treating the patient with an immunosuppressive agent.
9. The method of claim 7 wherein said immunosuppressive agent prevents an immune response.
10. The method of claim 7 wherein said immunosuppressive agent delays the occurrence of an immune response.
11. The method of claim 7 wherein said immunosuppressive agent decreases the intensity of an immune response.
12. The method of claim 7 or 8 , wherein said immune response is transplant rejection.
13. The method of claim 7 or 8 , wherein the immunosuppressive agent is selected from the group consisting of FK-506, cyclosporin, and GAD65 antibodies.
14. The method of claim 8 wherein said immunosuppressive agent prevents an immune response.
15. The method of claim 8 wherein said immunosuppressive agent delays the occurrence of an immune response.
16. The method of claim 8 wherein said immunosuppressive agent decreases the intensity of an immune response.
17. The method of claim 14 , 15 or 16 wherein said immune response is transplant rejection.