Differentiation of non-insulin producing cells into insulin producing cells by GLP-1 or exendin-4 and uses thereof
The present invention relates to a population of insulin producing cells made by a process comprising contacting non-insulin producing cells with a growth factor selected from the group consisting of GLP-1 or Exendin-4, growth factors having amino acid sequences substantially homologous to GLP-1 or Exendin-4, and fragmets thereof. The present invention also relates to methods of differentiating non-insulin producing cells into insulin producing cells and of enriching a population of cells for insulin-producing cells. The present invention also relates to methods of treating diabetes.
1. An isolated population of insulin-producing cells obtained from non-insulin-producing cells by a process comprising contacting the non-insulin-producing cells in vitro for at least twenty-four hours with an amount of a substance effective to induce insulin production, wherein the substance is selected from the group consisting of a GLP-1 peptide, a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1, and a fragment of any one of the preceding GLP-1 peptides, and wherein the GLP-1 peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells.
2. The population of claim 1 , wherein the non-insulin-producing cells comprise pancreatic cells.
3. The population of claim 1 , wherein the non-insulin-producing cells comprise pancreatic acinar cells.
4. The population of claim 1 , wherein the non-insulin-producing cells comprise stem cells.
5. The population of claim 1 , wherein the non-insulin-producing cells comprise pancreatic stem cells.
6. The population of claim 1 , wherein the non-insulin-producing cells comprise mammalian cells.
7. The population of claim 6 , wherein the mammalian cells comprise human cells.
8. The composition of claim 1 , wherein the contacting is at least 3 days.
9. The composition of claim 1 , wherein the substance is a GLP-1 peptide.
10. The composition of claim 1 , wherein the substance is a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1.
11. The composition of claim 1 , wherein the substance is the fragment of a GLP-1 peptide, or a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1.
12. A method for differentiating non-insulin-producing cells into insulin-producing cells, comprising contacting the non-insulin-producing cells in vitro for at least twenty four hours with an amount of a substance effective to induce differentiation of non-insulin-producing cells into insulin-producing cells, wherein the substance is selected from the group consisting of a GLP-1 peptide, a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1, and a fragment of any one of the preceding GLP-1 peptides, and wherein the GLP-1 peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells.
13. The method of claim 12 , wherein the non-insulin-producing cells comprise pancreatic cells, pancreatic acinar cells, stem cells, pancreatic stem cells, or mammalian cells.
14. The method of claim 13 , wherein the mammalian cells are human cells.
15. The method of claim 12 , wherein the contacting is at least 3 days.
16. The method of claim 12 , wherein the substance is a GLP-1 peptide.
17. The method of claims 12 , wherein the substance is a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1.
18. The method of claims 12 , wherein the substance is the fragment of a GLP-1 peptide, or a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1.
19. A method of enriching an isolated population of cells for insulin-producing cells, comprising contacting non-insulin-producing cells in vitro for at least twenty four hours with an amount of a substance effective to induce differentiation of non-insulin-producing cells into insulin-producing cells, wherein the substance is selected from the group consisting of a GLP-1 peptide, a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1, and a fragments of any one of the preceding peptides, and wherein the peptide or fragments thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells.
20. A method of promoting pancreatic amylase-producing cells to produce insulin, comprising contacting the pancreatic amylase-producing cells in vitro for at least twenty-four hours with an amount of a substance effective to induce insulin production, wherein the substance is selected from the group consisting of a GLP-1 peptide, a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1, and a fragment of any one of the preceding peptides, and wherein the GLP-1 peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells.
21. An isolated population of insulin-producing cells obtained from non-insulin-producing cells by a process comprising contacting the non-insulin-producing cells in vitro for at least twenty-four hours with an amount of a substance effective to induce insulin production, wherein the substance is selected from the group consisting of an Exendin-4 peptide, an Exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than positions 1, 4, 6, 7 and 9 of Exendin-4, and a fragment of any one of the preceding Exendin-4 peptides, and wherein the Exendin-4 peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells.
22. The population of claim 21 , wherein the non-insulin-producing cells comprise pancreatic cells.
23. The population of claim 21 , wherein the non-insulin-producing cells comprise pancreatic acinar cells.
24. The population of claim 21 , wherein the non-insulin-producing cells comprise stem cells.
25. The population of claim 21 , wherein the non-insulin-producing cells comprise pancreatic stem cells.
26. The population of claim 21 , wherein the non-insulin-producing cells comprise mammalian cells.
27. The population of claim 26 , wherein the mammalian cells comprise human cells.
28. The composition of claim 21 , wherein the contacting is at least 3 days.
29. The composition of claim 21 , wherein the substance is exendin-4.
30. The composition of claim 21 , wherein the substance is an Exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than positions 1, 4, 6, 7 and 9 of Exendin-4.
31. The composition of claim 21 , wherein the substance is the fragment of an Exendin-4 peptide, or an Exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than 1, 4, 6, 7 and 9 of Exendin-4.
32. A method for differentiating non-insulin-producing cells into insulin-producing cells, comprising contacting the non-insulin-producing cells in vitro for at least twenty four hours with an amount of a substance effective to induce differentiation of non-insulin-producing cells into insulin-producing cells, wherein the substance is selected from the group consisting of an Exendin-4 peptide, an Exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than positions 1, 4, 6, 7 and 9 of Exendin-4, and a fragment of any one of the preceding Exendin-4 peptides, and wherein the Exendin-4 peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells.
33. The method of claim 32 , wherein the non-insulin-producing cells comprise pancreatic cells, pancreatic acinar cells, stem cells, pancreatic stem cells, or mammalian cells.
34. The method of claim 33 , wherein the mammalian cells are human cells.
35. The method of claim 32 , wherein the contacting is at least 3 days.
36. The method of claim 32 , wherein the substance is exendin-4.
37. The method of claim 32 , wherein the substance is an Exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than positions 1, 4, 6, 7 and 9 of Exendin-4.
38. The method of claims 32 , wherein the substance is the fragment of an Exendin-4 peptide, or an Exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than 1, 4, 6, 7 and 9 of Exendin-4.
39. A method of promoting pancreatic amylase-producing cells to produce insulin, comprising contacting the pancreatic amylase-producing cells in vitro for at least twenty-four hours with an amount of a substance effective to induce insulin production, wherein the substance is selected from the group consisting of an Exendin-4 peptide, an Exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than 1, 4, 6, 7 and 9 of Exendin-4, and a fragment of any one of the preceding peptides, and wherein the Exendin-4 peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells.
40. A method of enriching an isolated population of cells for insulin-producing cells, comprising contacting non-insulin-producing cells in vitro for at least twenty-four hours with an amount of a substance effective to induce differentiation of non-insulin-producing cells into insulin-producing cells, wherein the substance is selected from the group consisting of an Exendin-4 peptide, an Exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than 1, 4, 6, 7 and 9 of Exendin-4, and a fragment of any one of the preceding peptides, and wherein the peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells.