Cofactors FAD and PLP for Promoting Pancreatic Progenitors in Pancreatic Development
The instant disclosure relates to methods and compositions for increasing β-cell production from stem cells.
1 . A method for producing endocrine pancreatic β-cells, the method comprising
contacting a culture including human induced pluripotent stem cells (hiPSCs) with at least one cofactor;
differentiating at least a portion of the hiPSCs to endocrine pancreatic β-cells; and
measuring expression of a biomarker selected from PDX1, NKX6, NGN3, or a combination thereof to identify the endocrine pancreatic β-cells, wherein expression of said biomarkers indicates differentiation to endocrine pancreatic β-cells.
2 . The method of claim 1 , wherein the hiPSCs are contacted with at least one cofactor during the pancreatic progenitor stage (S4) of differentiation of the hiPSCs.
3 . The method of claim 2 , wherein the at least one cofactor is contacted with the hiPSCs for a period of at least 4 days.
4 . The method of claim 1 , wherein the at least one cofactor is Flavin Adenine Dinucleotide (FAD), Pyridoxal 5′-phosphate (PLP), or a combination thereof.
5 . The method of claim 1 , wherein the at least one cofactor is FAD.
6 . The method of claim, wherein the at least one cofactor is PLP.
7 . The method of claim 1 , wherein the at least one cofactor is in a concentration range of about 0.1 μM to about 1.5 μM.
8 . The method of claim 1 , wherein the at least one cofactor is in a concentration range of about 0.25 μM to about 1 μM.
9 . The method of claim 1 , wherein the at least one cofactor is FAD at a concentration of about 0.5 μM.
10 . The method of claim 1 , wherein the at least one cofactor is in a concentration range of about 1 μM to about 10 μM.
11 . The method of claim 1 , wherein the at least one cofactor is in a concentration range of about 5 μM to about 10 μM.
12 . The method of claim 1 , wherein the at least one cofactor is PLP at a concentration of about 8 μM.
13 . The method of claim 1 , wherein the contacting of the culture including human induced pluripotent stem cells (hiPSCs) with at least one cofactor is conducted when the hiPSCs are about 70% confluent.
14 . A kit for use in producing endocrine pancreatic β-cells from a culture of human induced pluripotent stem cells (hiPSCs) comprising a composition comprising at least one cofactor.
15 . A method for treating diabetes in a subject comprising implanting a therapeutically effective amount of endocrine pancreatic β-cells, prepared according to the method of claim 1 , in the subject in need thereof.
16 . A method for regenerating β-cells in a subject, the method comprising implanting a therapeutically effective amount of endocrine pancreatic β-cells, prepared according to the method of claim 1 , in the subject in need thereof.
17 . A method for treating pancreatic cancer in a subject, the method comprising implanting a therapeutically effective amount of endocrine pancreatic β-cells, prepared according to the method of claim 1 , in the subject in need thereof.
18 . A composition comprising endocrine pancreatic β-cells and at least one cofactor.
19 . The composition of claim 18 , wherein the at least one cofactor is FAD, PLP, or a combination thereof.
20 . A therapeutic product prepared according to the method of claim 1 .