Method for manufacturing pancreatic-hormone-producing cells
First Claim
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1. A method of producing pancreatic hormone-producing cells, comprising:
- (i) culturing human or mouse induced-pluripotent stem (iPS) cells, in a medium containing an activator of activin receptor-like kinase-4,7 and a GSK3 inhibitor,(ii) culturing the cells of step (i) in a medium containing an activator of activin receptor-like kinase-4,7,(iii) culturing the cells obtained in step (ii) in a medium containing the following (a)-(c);
(a) retinoic acid receptor agonists,(b) at least one kind selected from the group consisting of inhibitors of AMP-activated protein kinase and/or activin receptor-like kinase-2,3,6, and BMP antagonists, and(c) inhibitors of activin receptor-like kinase-4,5,7, and(iv) culturing the cells obtained from step (iii) in a medium containing any factor selected from the group consisting of (ia) at least one kind selected from the group consisting of adenylate cyclase activators, cAMP phosphodiesterase inhibitors and cAMP analogs, (ib) steroids and (ic) inhibitors of activin receptor-like kinase-4,5,7 to obtain pancreatic hormone-producing cells.
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Abstract
The present invention provides a method of more efficiently producing pancreas cells, particularly pancreatic hormone-producing cells, a method of stably producing pancreas cells in a large amount by more efficiently inducing differentiation of stem cells into pancreas cells, a medicament containing a pancreas cells and a screening method using the cells.
A method of producing pancreatic hormone-producing cells, including subjecting stem cells to the following steps (1)-(4):
- (1) a step of cultivating stem cells in a medium containing an activator of activin receptor-like kinase-4,7 and a GSK3 inhibitor
- (2) a step of cultivating the cells obtained in the aforementioned step (1) in a medium containing an activator of activin receptor-like kinase-4,7
- (3) a step of cultivating the cells obtained in the aforementioned step (2) in a medium containing any one or more kinds selected from the group consisting of (a) retinoic acid receptor agonists, (b) at least one kind selected from the group consisting of inhibitors of AMP-activated protein kinase and/or activin receptor-like kinase-2,3,6, and BMP antagonists, and (c) inhibitors of activin receptor-like kinase-4,5,7
- (4) a step of cultivating the cells obtained in the aforementioned step (3).
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10 Claims
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1. A method of producing pancreatic hormone-producing cells, comprising:
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(i) culturing human or mouse induced-pluripotent stem (iPS) cells, in a medium containing an activator of activin receptor-like kinase-4,7 and a GSK3 inhibitor, (ii) culturing the cells of step (i) in a medium containing an activator of activin receptor-like kinase-4,7, (iii) culturing the cells obtained in step (ii) in a medium containing the following (a)-(c); (a) retinoic acid receptor agonists, (b) at least one kind selected from the group consisting of inhibitors of AMP-activated protein kinase and/or activin receptor-like kinase-2,3,6, and BMP antagonists, and (c) inhibitors of activin receptor-like kinase-4,5,7, and (iv) culturing the cells obtained from step (iii) in a medium containing any factor selected from the group consisting of (ia) at least one kind selected from the group consisting of adenylate cyclase activators, cAMP phosphodiesterase inhibitors and cAMP analogs, (ib) steroids and (ic) inhibitors of activin receptor-like kinase-4,5,7 to obtain pancreatic hormone-producing cells. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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Specification