Embryonic stem cells express neuronal properties in vitro

Dev Biol. 1995 Apr;168(2):342-57. doi: 10.1006/dbio.1995.1085.


Mouse embryonic stem (ES) cells cultured as aggregates and exposed to retinoic acid are induced to express multiple phenotypes normally associated with neurons. A large percentage of treated aggregates produce a rich neuritic outgrowth. Dissociating the induced aggregates with trypsin and plating the cells as a monolayer results in cultures in which a sizable percentage of the cells have a neuronal appearance. These neuron-like cells express class III beta-tubulin and the neurofilament M subunit. Induced cultures express transcripts for neural-associated genes including the neurofilament L subunit, glutamate receptor subunits, the transcription factor Brn-3, and GFAP. Levels of neurofilament L and GAD67 and GAD65 transcripts rise dramatically upon induction. Physiological studies show that the neuron-like cells generate action potentials and express TTX-sensitive sodium channels, as well as voltage-gated potassium channels and calcium channels. We conclude that a complex system of neuronal gene expression can be activated in cultured ES cells. This system should be favorable for investigating some of the mechanisms that regulate neuronal differentiation.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Cell Differentiation / drug effects*
  • Cells, Cultured
  • DNA, Complementary
  • Embryo, Mammalian / cytology
  • Membrane Potentials
  • Mice
  • Molecular Sequence Data
  • Nerve Tissue Proteins / biosynthesis
  • Neurons / cytology*
  • Neurons / physiology
  • Stem Cells / cytology*
  • Stem Cells / drug effects
  • Stem Cells / physiology
  • Tretinoin / pharmacology*
  • Tubulin / biosynthesis


  • DNA, Complementary
  • Nerve Tissue Proteins
  • Tubulin
  • Tretinoin