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 population of induced pluripotent stem cells, wherein said method comprises culturing said induced pluripotent stem cells with medium comprising indolactam V and glucagon like peptide-1 under conditions 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 medium lacks feeder cells.
4 . The method of claim 1 , wherein said medium lacks non-human feeder cells.
5 . The method of claim 1 , wherein said induced pluripotent stem cells were obtained using one or more polypeptides or nucleic acid encoding said one or more polypeptides 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.
6 . The method of claim 1 , wherein said induced pluripotent stem cells were induced from somatic cells.
7 . The method of claim 6 , wherein said somatic cells are selected from the group consisting of skin, lung, heart, stomach, brain, liver, blood, kidney, and muscle cells.
8 . 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.
9 . The method of claim 1 , wherein said glucose-responsive, insulin-secreting cells secrete greater than 50 pM of C peptide per hour when in culture in the presence of about 10 mM of glucose.
10 . The method of claim 1 , wherein said glucose-responsive, insulin-secreting cells secrete greater than 200 pM of C peptide per hour when in culture in the presence of about 10 mM of glucose.
11 . The method of claim 1 , wherein said glucose-responsive, insulin-secreting cells secrete between about 50 and 250 pM of C peptide per hour when in culture in the presence of about 10 mM of glucose.
12 . The method of claim 1 , wherein said glucose-responsive, insulin-secreting cells are human cells.
13 . The method of claim 1 , wherein said medium comprises greater than 300 nM of indolactam V.
14 . The method of claim 1 , wherein said medium comprises greater than 55 nM of glucagon like peptide-1.
15 . The method of claim 1 , wherein said culturing is performed for more than 25 days.
16 . A population of glucose-responsive, insulin-secreting cells derived from induced pluripotent stem cells, wherein said glucose-responsive, insulin-secreting cells are produced by culturing said induced pluripotent stem cells with medium comprising indolactam V and glucagon like peptide-1 under conditions that result in the formation of said population of glucose-responsive, insulin-secreting cells.
17 . The population of claim 15 , wherein said medium comprises greater than 300 nM of indolactam V.
18 . The population of claim 15 , wherein said medium comprises greater than 55 nM of glucagon like peptide-1.
19 . The population of claim 15 , wherein said culturing is performed for more than 25 days.
20 . The population of claim 15 , wherein said population of glucose-responsive, insulin-secreting cells secretes greater than 50 pM of C peptide per hour when in culture in the presence of about 10 mM of glucose.
21 . The population of claim 15 , wherein said population of glucose-responsive, insulin-secreting cells secrete greater than 200 pM of C peptide per hour when in culture in the presence of about 10 mM of glucose.
22 . The population of claim 15 , wherein said population of glucose-responsive, insulin-secreting cells secrete between about 50 and 250 pM of C peptide per hour when in culture in the presence of about 10 mM of glucose.
23 . The population of claim 15 , wherein said glucose-responsive, insulin-secreting cells are human cells.