Efficient generation of dopaminergic induced neuronal cells with midbrain characteristics

Stem Cell Reports. 2021 Jul 13;16(7):1763-1776. doi: 10.1016/j.stemcr.2021.05.017. Epub 2021 Jun 24.


The differentiation of pluripotent stem cells can be accomplished by sequential activation of signaling pathways or through transcription factor programming. Multistep differentiation imitates embryonic development to obtain authentic cell types, but it suffers from asynchronous differentiation with variable efficiency. Transcription factor programming induces synchronous and efficient differentiation with higher reproducibility but may not always yield authentic cell types. We systematically explored the generation of dopaminergic induced neuronal cells from mouse and human pluripotent stem cells. We found that the proneural factor Ascl1 in combination with mesencephalic factors Lmx1a and Nurr1 induce peripheral dopaminergic neurons. Co-delivery of additional midbrain transcription factors En1, FoxA2, and Pitx3 resulted in facile and robust generation of functional dopaminergic neurons of midbrain character. Our results suggest that more complex combinations of transcription factors may be needed for proper regional specification of induced neuronal cells generated by direct lineage induction.

Keywords: dopamine neurons; human embryonic stem cells; iN cells; mouse embyronic stem cells; reprogramming; transcription factor.

Publication types

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

MeSH terms

  • Animals
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Biomarkers / metabolism
  • Brain-Derived Neurotrophic Factor / metabolism
  • Cell Culture Techniques*
  • Dopamine / metabolism
  • Dopaminergic Neurons / cytology*
  • Embryonic Stem Cells / metabolism
  • Glial Cell Line-Derived Neurotrophic Factor / metabolism
  • Humans
  • Mesencephalon / cytology*
  • Mice
  • Signal Transduction
  • Transcription Factors / metabolism
  • Tyrosine 3-Monooxygenase / metabolism
  • Wnt1 Protein / metabolism


  • ASCL1 protein, human
  • Ascl1 protein, mouse
  • Basic Helix-Loop-Helix Transcription Factors
  • Biomarkers
  • Brain-Derived Neurotrophic Factor
  • Glial Cell Line-Derived Neurotrophic Factor
  • Transcription Factors
  • Wnt1 Protein
  • Tyrosine 3-Monooxygenase
  • Dopamine