Expression of early developmental markers predicts the efficiency of embryonic stem cell differentiation into midbrain dopaminergic neurons

Stem Cells Dev. 2013 Feb 1;22(3):397-411. doi: 10.1089/scd.2012.0238. Epub 2012 Sep 20.


Dopaminergic neurons derived from pluripotent stem cells are among the best investigated products of in vitro stem cell differentiation owing to their potential use for neurorestorative therapy of Parkinson's disease. However, the classical differentiation protocols for both mouse and human pluripotent stem cells generate a limited percentage of dopaminergic neurons and yield a considerable cellular heterogeneity comprising numerous scarcely characterized cell populations. To improve pluripotent stem cell differentiation protocols for midbrain dopaminergic neurons, we established extensive and strictly quantitative gene expression profiles, including markers for pluripotent cells, neural progenitors, non-neural cells, pan-neuronal and glial cells, neurotransmitter phenotypes, midbrain and nonmidbrain populations, floor plate and basal plate populations, as well as for Hedgehog, Fgf, and Wnt signaling pathways. The profiles were applied to discrete stages of in vitro differentiation of mouse embryonic stem cells toward the dopaminergic lineage and after transplantation into the striatum of 6-hydroxy-dopamine-lesioned rats. The comparison of gene expression in vitro with stages in the developing ventral midbrain between embryonic day 11.5 and 13.5 ex vivo revealed dynamic changes in the expression of transcription factors and signaling molecules. Based on these profiles, we propose quantitative gene expression milestones that predict the efficiency of dopaminergic differentiation achieved at the end point of the protocol, already at earlier stages of differentiation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Differentiation / genetics
  • Antigens, Differentiation / metabolism
  • Cell Differentiation*
  • Cells, Cultured
  • Dopaminergic Neurons / metabolism*
  • Dopaminergic Neurons / physiology
  • Dopaminergic Neurons / transplantation
  • Embryonic Stem Cells / metabolism*
  • Embryonic Stem Cells / physiology
  • Gene Expression
  • Genes, Developmental
  • Intracellular Signaling Peptides and Proteins / genetics
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Male
  • Mesencephalon / metabolism*
  • Mesencephalon / pathology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Neural Stem Cells / metabolism*
  • Neural Stem Cells / physiology
  • Neural Stem Cells / transplantation
  • Parkinson Disease, Secondary / pathology
  • Parkinson Disease, Secondary / therapy
  • Rats
  • Rats, Wistar
  • Signal Transduction
  • Transcriptome


  • Antigens, Differentiation
  • Intracellular Signaling Peptides and Proteins