GENERATION OF PANCREATIC ENDOCRINE CELLS FROM PRIMARY DUCT CELL CULTURES AND METHODS OF USE FOR TREATMENT OF DIABETES
First Claim
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1. An immortalized pancreatic duct cell derived from a primary adult pancreatic duct epithelial cell culture, wherein the immortalized cell expresses Pdx1 and FoxO1.
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Abstract
The invention is directed to spontaneously immortalized pancreatic duct cells and methods for generating pancreatic endocrine cells from spontaneously immortalized pancreatic duct cells that express the transcription factors Pdx1 and FoxO1. The invention also provides for methods for treating beta cell failure, the method comprising administering to a subject an effective amount of spontaneously immortalized pancreatic duct cells expressing a mutated version FoxO1.
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17 Claims
- 1. An immortalized pancreatic duct cell derived from a primary adult pancreatic duct epithelial cell culture, wherein the immortalized cell expresses Pdx1 and FoxO1.
- 9. An immortalized pancreatic duct cell derived from a primary adult pancreatic duct epithelial cell, wherein the cell expresses mutated FoxO1.
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13. A pancreatic duct cell line designated 24-1 Duct having ATCC Accession No. PTA-6968.
- 14. A method for treating beta cell failure, the method comprising administering to a subject spontaneously immortalized pancreatic ductal cell line expressing mutated FoxO1.
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17. A method for obtaining a pancreatic duct cell line, the method comprising:
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(a) culturing pancreatic duct cells collected from a subject in medium comprising about 10% serum and 5.5 mM glucose for about a week; (b) culturing the cells in a medium comprising (i) about 8 mM glucose;
(ii) about 1 g/L ITS (about 5 mg/l insulin and about 5 mg/l transferrin and about 5 mg/l selectin), (iii) about 2 g/l albumin, (iv) about 10 mM nicotinamide, and (v) about 10 mg/ml keratinocyte growth factor, for about at least another week until the culture comprises nearly all duct cells;(c) culturing the duct cells with the medium of step (b) further comprising about 10% serum and about 5.5 mM glucose; (d) passaging the cells of step (c) until the cells'"'"' doubling time reach about 24 hours; and (e) cloning a single cell from the cells of step (d) so as to obtain a pancreatic duct cell line.
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Specification