Islet cells from human embryonic stem cells
This disclosure provides a system for producing pancreatic islet cells from embryonic stem cells. Differentiation is initiated towards endoderm cells, and focused using reagents that promote emergence of islet precursors and mature insulin-secreting cells. High quality populations of islet cells can be produced in commercial quantities for use in research, drug screening, or regenerative medicine.
1. A method for obtaining human insulin secreting cells, comprising
a) culturing hES cells in a first medium comprising Activin A; and
b) maturing the cells from a) in a second medium comprising nicotinamide; thereby obtaining a population comprising human insulin secreting cells.
2. The method of claim 1 , wherein the first medium further comprises n-butyrate.
3. A method for obtaining human insulin secreting cells, comprising
a) culturing hES cells in a suspension culture to form embryoid bodies;
b) culturing the cells from a) in a medium comprising trans-retinoic acid;
c) culturing the cells from b) in a medium comprising a TGF-β antagonist and one or more mitogens; and
d) maturing the cells from c) in a medium comprising nicotinamide; thereby obtaining a population comprising human insulin-secreting cells.
4. The method of claim 3 , wherein the TGF-β antagonist is Noggin.
5. The method of claim 3 , wherein the mitogen(s) is selected from the group consisting of EGE, bFGF, and betacellulin.
6. A method for obtaining human insulin secreting cells, comprising:
a) culturing hES cells in a first medium comprising Activin A;
b) culturing the cells from step a) in a second medium comprising a TGF-β antagonist and one or more mitogens; and
c) culturing the cells from step b) in a third medium comprising nicotinamide; thereby obtaining a population comprising human insulin-secreting cells.
7. The method of claim 6 , wherein the TGF-β antagonist is Noggin.
8. The method of claim 6 , wherein the mitogen(s) is selected from the group consisting of epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), and betacellulin.
9. The method of claim 6 , wherein the second medium comprises Noggin, EGF, and bFGF.
10. The method of claim 6 , wherein the second medium comprises Noggin, betacellulin, and bFGF.