title: Rapid generation of functional dopaminergic neurons from human induced pluripotent stem cells through a single-step procedure using cell lineage transcription factors.
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.
authors: Division of Neuroscience, San Raffaele Scientific Institute, Milan, Italy.
authors: Ilda
authors: I
authors: Theka
authors: Massimiliano
authors: M
authors: Caiazzo
authors: Elena
authors: E
authors: Dvoretskova
authors: Damiana
authors: D
authors: Leo
authors: Federica
authors: F
authors: Ungaro
authors: Sebastiano
authors: S
authors: Curreli
authors: Francesca
authors: F
authors: Managò
authors: Maria Teresa
authors: MT
authors: Dell'Anno
authors: Gianni
authors: G
authors: Pezzoli
authors: Raul R
authors: RR
authors: Gainetdinov
authors: Alexander
authors: A
authors: Dityatev
authors: Vania
authors: V
authors: Broccoli
mesh_terms: D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D019070:Cell Lineage / Q000502:physiology*; D002478:Cells, Cultured; D004298:Dopamine / Q000378:metabolism*; D059290:Dopaminergic Neurons / Q000166:cytology* / Q000502:physiology*; D018507:Gene Expression Regulation, Developmental; D005822:Genetic Vectors; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology; D060850:LIM-Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics; D008564:Membrane Potentials / Q000502:physiology; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2 / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014404:Tubulin / Q000235:genetics / Q000378:metabolism
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D015415:Biomarkers; D004298:Dopamine; D060850:LIM-Homeodomain Proteins; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D014157:Transcription Factors; D014404:Tubulin; C065500:ASCL1 protein, human; C078021:LMX1A protein, human; C539882:NR4A2 protein, human; C498585:TUBB3 protein, human
keywords: Direct cell conversion; Dopamine; Neuron; Pluripotent stem cells; Reprogramming
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