Differentiating induced pluripotent stem cells into glucose-responsive, insulin-secreting progeny
This document provides methods and materials related to differentiating iPS cells into glucose-responsive, insulin-secreting progeny. For example, methods and material for using indolactam V (ILV) and glucagon like peptide-1 (GLP-1) to produce glucose-responsive, insulin-secreting progeny from iPS cells are provided.
1. A method for obtaining a population of glucose-responsive, insulin-secreting cells from a diabetic human, wherein said method comprises:
(a) obtaining somatic cells from a diabetic human,
(b) exposing said somatic cells to one or more polypeptides or nucleic acids encoding said one or more polypeptides to form induced pluripotent stem cells, wherein said one or more polypeptides are selected from the group consisting of a Oct3/4 polypeptide, a Sox family polypeptide, a Klf family polypeptide, a Myc family polypeptide, a Nanog polypeptide, and a Lin28 polypeptide, and
(c) culturing said induced pluripotent stem cells with medium comprising indolactam V and glucagon like peptide-1 to obtain said population of glucose-responsive, insulin-secreting cells.
2. The method of claim 1 , wherein said medium lacks serum.
3. The method of claim 1 , wherein said culturing is performed in the absence of feeder cells.
4. The method of claim 1 , wherein said culturing is performed in the absence of non-human feeder cells.
5. The method of claim 1 , wherein said somatic cells are selected from the group consisting of skin, lung, heart, stomach, brain, liver, blood, kidney, and muscle cells.
6. The method of claim 1 , wherein said induced pluripotent stem cells comprise exogenous nucleic acid encoding a human Oct4 polypeptide, a human Sox2 polypeptide, a human Klf4 polypeptide, and a human c-Myc polypeptide.
7. The method of claim 1 , wherein said medium comprises greater than 300 nM of indolactam V.
8. The method of claim 1 , wherein said medium comprises greater than 55 nM of glucagon like peptide-1.
9. The method of claim 1 , wherein said culturing is performed for four days.