A Modular Platform for Differentiation of Human PSCs into All Major Ectodermal Lineages

Cell Stem Cell. 2017 Sep 7;21(3):399-410.e7. doi: 10.1016/j.stem.2017.08.015.


Directing the fate of human pluripotent stem cells (hPSCs) into different lineages requires variable starting conditions and components with undefined activities, introducing inconsistencies that confound reproducibility and assessment of specific perturbations. Here we introduce a simple, modular protocol for deriving the four main ectodermal lineages from hPSCs. By precisely varying FGF, BMP, WNT, and TGFβ pathway activity in a minimal, chemically defined medium, we show parallel, robust, and reproducible derivation of neuroectoderm, neural crest (NC), cranial placode (CP), and non-neural ectoderm in multiple hPSC lines, on different substrates independently of cell density. We highlight the utility of this system by interrogating the role of TFAP2 transcription factors in ectodermal differentiation, revealing the importance of TFAP2A in NC and CP specification, and performing a small-molecule screen that identified compounds that further enhance CP differentiation. This platform provides a simple stage for systematic derivation of the entire range of ectodermal cell types.

Keywords: TFAP2A; chemical screen; chemically defined Ectoderm; cortical neurons; cranial placode; directed differentiation; human pluripotent stem cells; neural crest; non-neural ectoderm.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Bone Morphogenetic Proteins / metabolism
  • Cell Differentiation* / drug effects
  • Cell Lineage* / drug effects
  • Ectoderm / cytology*
  • Gene Expression Regulation, Developmental / drug effects
  • Humans
  • Neural Crest / cytology
  • Neural Plate / cytology
  • Neural Stem Cells / cytology
  • Neural Stem Cells / drug effects
  • Neural Stem Cells / metabolism
  • Phenanthrolines / pharmacology
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / drug effects
  • Pluripotent Stem Cells / metabolism
  • Signal Transduction / drug effects
  • Small Molecule Libraries / pharmacology
  • Transcription Factor AP-2 / metabolism


  • Bone Morphogenetic Proteins
  • Phenanthrolines
  • Small Molecule Libraries
  • TFAP2A protein, human
  • Transcription Factor AP-2