Open AccessRapid Generation of Functional Dopaminergic Neurons From Human Induced Pluripotent Stem Cells Through a Single‐Step Procedure Using Cell Lineage Transcription Factors
Author(s) -
Theka Ilda,
Caiazzo Massimiliano,
Dvoretskova Elena,
Leo Damiana,
Ungaro Federica,
Curreli Sebastiano,
Managò Francesca,
Dell'Anno Maria Teresa,
Pezzoli Gianni,
Gainetdinov Raul R.,
Dityatev Alexander,
Broccoli Vania
Publication year - 2013
Publication title -
stem cells translational medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.781
H-Index - 71
eISSN - 2157-6580
pISSN - 2157-6564
DOI - 10.5966/sctm.2012-0133
Subject(s) - induced pluripotent stem cell , embryoid body , biology , dopaminergic , transcription factor , dopamine , stem cell , directed differentiation , cellular differentiation , embryonic stem cell , neuroscience , microbiology and biotechnology , genetics , gene
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.