Efficient endoderm induction from human pluripotent stem cells by logically directing signals controlling lineage bifurcations

Cell Stem Cell. 2014 Feb 6;14(2):237-52. doi: 10.1016/j.stem.2013.12.007. Epub 2014 Jan 9.


Human pluripotent stem cell (hPSC) differentiation typically yields heterogeneous populations. Knowledge of signals controlling embryonic lineage bifurcations could efficiently yield desired cell types through exclusion of alternate fates. Therefore, we revisited signals driving induction and anterior-posterior patterning of definitive endoderm to generate a coherent roadmap for endoderm differentiation. With striking temporal dynamics, BMP and Wnt initially specified anterior primitive streak (progenitor to endoderm), yet, 24 hr later, suppressed endoderm and induced mesoderm. At lineage bifurcations, cross-repressive signals separated mutually exclusive fates; TGF-β and BMP/MAPK respectively induced pancreas versus liver from endoderm by suppressing the alternate lineage. We systematically blockaded alternate fates throughout multiple consecutive bifurcations, thereby efficiently differentiating multiple hPSC lines exclusively into endoderm and its derivatives. Comprehensive transcriptional and chromatin mapping of highly pure endodermal populations revealed that endodermal enhancers existed in a surprising diversity of "pre-enhancer" states before activation, reflecting the establishment of a permissive chromatin landscape as a prelude to differentiation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Body Patterning / drug effects
  • Bone Morphogenetic Proteins / metabolism
  • Cell Lineage* / drug effects
  • Chromatin / metabolism
  • Culture Media, Serum-Free / pharmacology
  • Digestive System / embryology
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / metabolism
  • Endoderm / cytology
  • Endoderm / drug effects
  • Endoderm / embryology*
  • Enhancer Elements, Genetic / genetics
  • Epigenesis, Genetic / drug effects
  • Fibroblast Growth Factors / metabolism
  • Humans
  • Liver / embryology
  • MAP Kinase Signaling System / drug effects
  • Mice
  • Molecular Sequence Data
  • Pancreas / embryology
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / drug effects
  • Primitive Streak / cytology
  • Primitive Streak / embryology
  • Protein Binding / drug effects
  • Signal Transduction* / drug effects
  • Transcription, Genetic / drug effects
  • Transforming Growth Factor beta / metabolism
  • Wnt Proteins / metabolism


  • Bone Morphogenetic Proteins
  • Chromatin
  • Culture Media, Serum-Free
  • Transforming Growth Factor beta
  • Wnt Proteins
  • Fibroblast Growth Factors

Associated data

  • GEO/GSE52658