SC-BETA CELLS AND COMPOSITIONS AND METHODS FOR GENERATING THE SAME
Disclosed herein are methods, compositions, kits, and agents useful for inducing β cell maturation, and isolated populations of SC-β cells for use in various applications, such as cell therapy.
1 . A method of differentiating progenitor cells into pancreatic β cells in vitro, the method comprising culturing a cluster of the progenitor cells in suspension in a culture medium comprising the following compounds: a thyroid hormone signaling pathway activator and a transforming growth factor β (TGF-β) signaling pathway inhibitor, thereby differentiating the progenitor cells into pancreatic β cells, wherein the pancreatic β cells exhibit an in vitro glucose-stimulated insulin secretion response to at least a first glucose challenge and a second glucose challenge when the first glucose challenge and the second glucose challenge are applied sequentially.
2 . The method of claim 1 , wherein the progenitor cells comprise an insulin-positive endocrine cell.
3 . The method of claim 1 , wherein the progenitor cells comprise a PDX1-positive progenitor cell.
4 . The method of claim 1 , wherein the thyroid hormone signaling pathway activator is triiodothyronine.
5 . The method of claim 4 , wherein the concentration of the triiodothyronine is at least 0.1 μM.
6 . The method of claim 1 , wherein the TGF-β signaling pathway inhibitor is Alk5 inhibitor II.
7 . The method of claim 6 , wherein the concentration of the Alk5 inhibitor II is at least 100 nM.
8 . The method of claim 1 , wherein the culturing comprises changing the culture medium at least every other day.
9 . The method of claim 1 , wherein the culturing lasts for at least seven days.
10 . The method of claim 1 , wherein the culture medium is a serum-free culture medium.
11 . The method of claim 1 , wherein the progenitor cells are cultured in the presence of a low-adherent substrate.
12 . The method of claim 1 , wherein the progenitor cells are human progenitor cells.
13 . The method of claim 1 , wherein the progenitor cells are generated from stem cells.
14 . The method of claim 13 , wherein the stem cells are embryonic stem cells.
15 . The method of claim 13 , wherein the stem cells are induced pluripotent stem cells.
16 . The method of claim 1 , wherein the non-native pancreatic β cells exhibit an in vitro glucose-stimulated insulin secretion response to a first glucose challenge, a second glucose challenge, and a third glucose challenge, when the first glucose challenge, the second glucose challenge and the third glucose challenge are applied sequentially.