Differentiating induced pluripotent stem cells into glucose-responsive, insulin-secreting progeny

Abstract

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.