Claims
- 1. A method for transforming IP cells that express markers of acinar cells and liver-associated genes into insulin-producing cells in vitro, comprising culturing said IP cells in a cell culture medium comprising an effective amount of at least one differentiation promoting factor selected from the group consisting of C-Natriuretic Peptide (CNP), Calcitonin Gene Related Peptide, Cholera Toxin B Subunit, Dexamethasone, Gastrin-Releasing Peptide, Laminin, Met-Enkephalin, PDGFAA+PDGFBB, Sonic Hedgehog, and Substance P such that the IP cells are transformed into insulin-producing cells.
- 2. The method of claim 1, wherein the IP cells are derived from a culture of pancreatic acinar cells.
- 3. The method of claim 2, wherein the cells are human.
- 4. The method of claim 1, further comprising contacting said cells with a substrate that is coated with one or more extracellular matrix molecules.
- 5. The method of claim 4, wherein the extracellular matrix molecules are collagen I, collagen VI, collagen IV, vitronectin, and/or fibronectin.
- 6. The method of claim 4, wherein the substrate is on the surface of a flask, petri dish, plate, well or roller bottle, or is part of a scaffold.
- 7. The method of claim 1, wherein the medium is serum-free.
- 8. The method of claim 1, wherein the medium comprises serum.
- 9. The method of claim 7, wherein the medium comprises BSA, insulin, transferrin, selenium and epidermal growth factor (EGF).
- 10. The method of claim 3, wherein the cells are seeded on the substrate at a density of 5×103 to 20×105 cells/cm2.
- 11. An isolated insulin-producing cell generated by the method of claim 1.
- 12. An insulin-producing cell, prepared by differentiating a mammalian acinar cell in vitro, wherein said insulin-producing cell has an expression profile after 16 days ex vivo as shown in Table 6.
- 13. A serum-free medium comprising at least one differentiation promoting factor selected from the group consisting of C-Natriuretic Peptide (CNP), Calcitonin Gene Related Peptide, Cholera Toxin B Subunit, Dexamethasone, Gastrin-Releasing Peptide, Laminin, Met-Enkephalin, PDGFAA+PDGFBB, Sonic Hedgehog, and Substance P wherein said medium facilitates differentiation of IP cells into insulin-producing cells.
- 14. A serum free medium comprising a 1:1 mixture of DMEM and Hams F12 plus the components listed in Table 2.
- 15. A kit suitable for differentiating IP cells to insulin-producing cells, comprising
a) a base medium suitable for the cultivation of mammalian epithelial cells; b) a collagen I coated culture substrate, and, separately packaged, c) a serum-free medium supplement containing BSA, C-Natriuretic Peptide (CNP), Calcitonin Gene Related Peptide, Cholera Toxin B Subunit, Dexamethasone, Gastrin-Releasing Peptide, Laminin, Met-Enkephalin, PDGFAA+PDGFBB, Sonic Hedgehog, and Substance P or two or more of these components in combination, in suitable amounts to yield final concentrations in the completed medium as indicated in Table 1 herein.
- 16. The kit of claim 15, wherein the cell culture substrate is contained on the surface of a flask, bottle, petri dish, plate or well suitable for cell culture.
- 17. The method of claim 1, wherein the cell culture medium comprises a 1: 1 mixture of DMEM and Hams 12.
- 18. The serum-free medium of claim 13 which comprises a 1:1 mixture of DMEM and Hams 12.
- 19. The method of claim 1, wherein said differentiation promoting factors have the concentrations in the medium as indicated in Table 1.
- 20. The serum-free medium of claim 13, wherein said differentiation promoting factors have the concentrations in the medium as indicated in Table 1.
Parent Case Info
[0001] This application claims the benefit of provisional application 60/384,000, filed May 28, 2002, which disclosure is incorporated by reference in its entirety herein.
Provisional Applications (1)
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Number |
Date |
Country |
|
60384000 |
May 2002 |
US |