Compositions and methods for promoting the generation of definitive endoderm

摘要

Certain embodiments disclosed herein are directed to a method of producing endoderm cells, such as definitive endoderm cells by exposing stem cells such as embryonic stem cells or induced pluripotent stem (iPS) cells to an effective amount of at least one compound described herein to differentiate the stem cells into the endoderm cells such as definitive endoderm cells. Differentiated endoderm cells produced by the methods disclosed herein can be differentiated into pancreatic epithelium, and other endoderm derivatives such as thymus, liver, stomach, intestine and lung. Another aspect of the present invention relates to a method of producing pancreatic progenitor cells, such as Pdx1-positive pancreatic progenitor cells by exposing endoderm cells, such as definitive endoderm cells to an effective amount of at least one compound described herein to differentiate the definitive endoderm cells into Pdx1-positive pancreatic progenitor cells. Kits and compositions comprising Pdx1-positive pancreatic progenitor produced using the methods are also described.

主权项

1. A method comprising:
(a) contacting a population of pluripotent stem cells with at least one compound of Formula (I) that activates TGF-β signaling, thereby inducing the differentiation of at least one pluripotent stem cell into a definitive endoderm cell, wherein the definitive endoderm cell expresses Sox17, or HNF3B (FoxA2), or Sox17 and HNF3B (FoxA2) and wherein the compound of formula (I) is: wherein: R1 and R2 are independently H, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cyclyl, or cyclyl, each of which can be optionally substituted and/or can be interrupted in the backbone with one or more of O, N, S, S(O), and C(O); R3 and R4 are independently H, halogen, alkyl, alkenyl, alkynyl, alkoxy, aryl, heteroaryl, cyclyl, or cyclyl, each of which can be optionally substituted, or R3 and R4 together with the carbon to which they are attached from an optionally substituted cyclyl or heterocyclyl; and L is C1-C10 alkylenyl, C2-C10 alkenylenyl, or C2-C10 alkynylenyl, each of which can be optionally substituted and/or can be interrupted in the backbone with one or more of O, N, S, S(O), and C(O), thereby producing a definitive endoderm cell from a pluripotent stem cell; and (b) differentiating the definitive endoderm cell into a Pdx1-positive pancreatic progenitor cell, wherein the Pdx1-positive pancreatic progenitor cell expresses Pdx1.