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

Autor: Damiana Leo, Alexander Dityatev, Raul R. Gainetdinov, Federica Ungaro, Maria Teresa Dell’Anno, Vania Broccoli, Francesca Managò, Massimiliano Caiazzo, Ilda Theka, Gianni Pezzoli, Sebastiano Curreli, Elena Dvoretskova
Přispěvatelé: 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
Jazyk: angličtina
Rok vydání: 2013
Předmět:
Transcription Factor
Dopamine
Cellular differentiation
Genetic Vectors
Induced Pluripotent Stem Cells
LIM-Homeodomain Proteins
Pluripotent stem cell
Basic Helix-Loop-Helix Transcription Factor
Embryoid body
Biology
LIM-Homeodomain Protein
Membrane Potential
Induced Pluripotent Stem Cell
Lentiviru
Membrane Potentials
Tubulin
Tissue Engineering and Regenerative Medicine
Nuclear Receptor Subfamily 4
Group A
Member 2
Basic Helix-Loop-Helix Transcription Factors
medicine
Humans
Cell Lineage
Induced pluripotent stem cell
Transcription factor
Cells
Cultured
Dopaminergic Neurons
Lentivirus
Dopaminergic
Gene Expression Regulation
Developmental
Reprogramming
Cell Differentiation
Biomarker
Cell Biology
General Medicine
Neuron
Molecular biology
Cell biology
ASCL1
medicine.anatomical_structure
Direct cell conversion
Genetic Vector
Dopaminergic Neuron
Biomarkers
Transcription Factors
Human
Developmental Biology
Popis: 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.
Databáze: OpenAIRE
Externí odkaz: