Rapid generation of functional dopaminergic neurons from human induced pluripotent stem cells through a single-step procedure using cell lineage transcription factors

Ilda Theka, Massimiliano Caiazzo, Elena Dvoretskova, Damiana Leo, Federica Ungaro, Sebastiano Curreli, Francesca Managò, Maria Teresa Dell'Anno, Gianni Pezzoli, Raul R. Gainetdinov, Alexander Dityatev, Vania Broccoli

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Current protocols for in vitro differentiation of human induced pluripotent stem cells (hiPSCs) to generate dopamine (DA) neurons are laborious and time-expensive. In order to accelerate the overall process, we have established a fast protocol by expressing the developmental transcription factors ASCL1, NURR1, and LMX1A. With this method, we were able to generate mature and functional dopaminergic neurons in as few as 21 days, skipping all the intermediate steps for inducting and selecting embryoid bodies and rosette-neural precursors. Strikingly, the resulting neuronal conversion process was very proficient, with an overall efficiency that was more than 93% of all the coinfected cells. hiPSC-derived DA neurons expressed all the critical molecular markers of the DA molecular machinery and exhibited sophisticated functional features including spontaneous electrical activity and dopamine release. This one-step protocol holds important implications for in vitro disease modeling and is particularly amenable for exploitation in high-throughput screening protocols.

Original languageEnglish
Pages (from-to)473-9
Number of pages7
JournalStem cells translational medicine
Volume2
Issue number6
DOIs
Publication statusPublished - Jun 2013

Keywords

  • Basic Helix-Loop-Helix Transcription Factors
  • Biomarkers
  • Cell Differentiation
  • Cell Lineage
  • Cells, Cultured
  • Dopamine
  • Dopaminergic Neurons
  • Gene Expression Regulation, Developmental
  • Genetic Vectors
  • Humans
  • Induced Pluripotent Stem Cells
  • LIM-Homeodomain Proteins
  • Lentivirus
  • Membrane Potentials
  • Nuclear Receptor Subfamily 4, Group A, Member 2
  • Transcription Factors
  • Tubulin
  • Journal Article
  • Research Support, Non-U.S. Gov't

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