Combined small-molecule inhibition accelerates developmental timing and converts human pluripotent stem cells into nociceptors

Nat Biotechnol. 2012 Jul 1;30(7):715-20. doi: 10.1038/nbt.2249.


Considerable progress has been made in identifying signaling pathways that direct the differentiation of human pluripotent stem cells (hPSCs) into specialized cell types, including neurons. However, differentiation of hPSCs with extrinsic factors is a slow, step-wise process, mimicking the protracted timing of human development. Using a small-molecule screen, we identified a combination of five small-molecule pathway inhibitors that yield hPSC-derived neurons at >75% efficiency within 10 d of differentiation. The resulting neurons express canonical markers and functional properties of human nociceptors, including tetrodotoxin (TTX)-resistant, SCN10A-dependent sodium currents and response to nociceptive stimuli such as ATP and capsaicin. Neuronal fate acquisition occurs about threefold faster than during in vivo development, suggesting that use of small-molecule pathway inhibitors could become a general strategy for accelerating developmental timing in vitro. The quick and high-efficiency derivation of nociceptors offers unprecedented access to this medically relevant cell type for studies of human pain.

Publication types

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

MeSH terms

  • Acetanilides / pharmacology
  • Caffeic Acids / pharmacology
  • Cell Culture Techniques
  • Cell Differentiation* / drug effects
  • Cell Line
  • Gene Expression Regulation, Developmental / drug effects
  • Humans
  • Molecular Sequence Data
  • NAV1.8 Voltage-Gated Sodium Channel / metabolism
  • Nociceptors* / cytology
  • Nociceptors* / drug effects
  • Nociceptors* / metabolism
  • Pain / metabolism
  • Pain / physiopathology
  • Pluripotent Stem Cells* / cytology
  • Pluripotent Stem Cells* / drug effects
  • Pluripotent Stem Cells* / metabolism
  • Pyridines / pharmacology
  • Pyrimidines / pharmacology
  • Pyrroles / pharmacology
  • Signal Transduction / drug effects
  • Small Molecule Libraries* / chemistry
  • Small Molecule Libraries* / pharmacology
  • Tetrodotoxin / pharmacology


  • Acetanilides
  • Caffeic Acids
  • Chir 99021
  • N-(4-(3-(3,4-dihydroxyphenyl)acryloyl)phenyl)-2-(thiophen-2-yl)acetamide
  • NAV1.8 Voltage-Gated Sodium Channel
  • Pyridines
  • Pyrimidines
  • Pyrroles
  • SCN10A protein, human
  • SU 5402
  • Small Molecule Libraries
  • Tetrodotoxin

Associated data

  • GENBANK/GSE26867