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 pluripotent stem cells in vitro with a first medium comprising an agent that activates a TGF-13 receptor family member;
(b) culturing in vitro the human pluripotent stem cell-derived cells of step (a) in a second medium lacking a substantial amount of the agent that activates 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.
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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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Citations
25 Claims
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1. A method for producing insulin, said method comprising the steps of:
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(a) contacting human pluripotent stem cells in vitro with a first medium comprising an agent that activates a TGF-13 receptor family member; (b) culturing in vitro the human pluripotent stem cell-derived cells of step (a) in a second medium lacking a substantial amount of the agent that activates 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 (2, 3, 4, 5, 6, 7, 8)
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9. A method for producing insulin, said method comprising the steps of:
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(a) contacting human pluripotent stem cells in vitro with a medium comprising a first agent that activates a TGF-β
receptor family member;(b) culturing in vitro the human pluripotent cell-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 (10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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20. A method for producing insulin, said method comprising the steps of:
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(a) contacting human pluripotent stem cells in vitro with a medium comprising an agent that activates a TGF-β
receptor family member;(b) removing the medium from the cell culture of step (a); (c) transplanting the human pluripotent stem cells-derived cells of step (b) into a mammalian subject; and (d) maturing the human pluripotent stem cells-derived cells of step (b) in vivo, thereby obtaining insulin secreting cells. - View Dependent Claims (21, 22, 23, 24, 25)
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Specification