Methods of producing pancreatic hormones
First Claim
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1. A method for producing insulin, said method comprising the steps of:
- (a) contacting human embryonic stem (hES) cells in vitro with a first medium comprising activin A, thereby activating a TGF-β
receptor family member;
(b) culturing in vitro the hES-derived cells of step (a) in a second medium lacking activin A, thereby generating PDX-1 positive pancreatic endoderm cells;
(c) transplanting the PDX-1 positive pancreatic endoderm cells of step (b) into a mammalian subject; and
(d) maturing the PDX-1 positive pancreatic endoderm cells of step (c) in vivo, thereby obtaining insulin secreting cells, wherein the insulin secreting cells secrete insulin in response to glucose stimulation.
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Abstract
Disclosed herein are methods of producing pancreatic hormone-expressing cells by first differentiating pluripotent cells in cell culture so as to produce endodermal cells, the endodermal cells being competent to further differentiate into hormone-expressing cells capable of secreting at least one pancreatic hormone in response to a physiological signal, and then, transplanting the cultured endodermal cells into an organism, such as an organism in need of an endocrine cell therapy.
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11 Claims
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1. A method for producing insulin, said method comprising the steps of:
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(a) contacting human embryonic stem (hES) cells in vitro with a first medium comprising activin A, thereby activating a TGF-β
receptor family member;(b) culturing in vitro the hES-derived cells of step (a) in a second medium lacking activin A, thereby generating PDX-1 positive pancreatic endoderm cells; (c) transplanting the PDX-1 positive pancreatic endoderm cells of step (b) into a mammalian subject; and (d) maturing the PDX-1 positive pancreatic endoderm cells of step (c) in vivo, thereby obtaining insulin secreting cells, wherein the insulin secreting cells secrete insulin in response to glucose stimulation. - View Dependent Claims (2, 3, 4, 5, 6)
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7. A method for producing insulin, said method comprising the steps of:
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(a) contacting human embryonic stem (hES) cells in vitro with a medium comprising activin A, thereby activating a TGF-β
receptor family member,(b) culturing in vitro the hES-derived cells of step (a) in a second medium comprising a second agent that inhibits the TGF-β
receptor family member, thereby generating PDX-1 positive pancreatic endoderm cells;(c) transplanting the PDX-1 positive pancreatic endoderm cells of step (b) into a mammalian subject; and (d) maturing the PDX-1 positive pancreatic endoderm cells of step (c) in vivo, thereby obtaining insulin secreting cells, wherein the insulin secreting cells secrete insulin in response to glucose stimulation. - View Dependent Claims (8, 9, 10, 11)
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Specification