Combining Suppression of Stemness with Lineage-Specific Induction Leads to Conversion of Pluripotent Cells into Functional Neurons

Chem Biol. 2015 Nov 19;22(11):1512-1520. doi: 10.1016/j.chembiol.2015.10.008.

Abstract

Sox2 is a key player in the maintenance of pluripotency and stemness, and thus inhibition of its function would abrogate the stemness of pluripotent cells and induce differentiation into several types of cells. Herein we describe a strategy that relies on a combination of Sox2 inhibition with lineage-specific induction to promote efficient and selective differentiation of pluripotent P19 cells into neurons. When P19 cells transduced with Skp protein, an inhibitor of Sox2, are incubated with a neurogenesis inducer, the cells are selectively converted into neurons that generate depolarization-induced sodium currents and action potentials. This finding indicates that the differentiated neurons are electrophysiologically active. Signaling pathway studies lead us to conclude that a combination of Skp with the neurogenesis inducer enhances neurogenesis in P19 cells by activating Wnt and Notch pathways. The present differentiation protocol could be valuable to selectively generate functionally active neurons from pluripotent cells.

Keywords: electrophysiology; neurogenesis; pluripotency; small molecules; stemness.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Cell Lineage
  • Mice
  • Microscopy, Fluorescence
  • Neurogenesis / drug effects
  • Neurons / cytology
  • Neurons / metabolism
  • Patch-Clamp Techniques
  • Peptides / chemistry
  • Peptides / pharmacology
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / metabolism
  • SOXB1 Transcription Factors / antagonists & inhibitors
  • SOXB1 Transcription Factors / metabolism*
  • Signal Transduction / drug effects
  • Small Molecule Libraries / chemistry
  • Small Molecule Libraries / pharmacology

Substances

  • Peptides
  • SOXB1 Transcription Factors
  • Small Molecule Libraries
  • Sox2 protein, mouse