title: Efficient induction of dopaminergic neuron differentiation from induced pluripotent stem cells reveals impaired mitophagy in PARK2 neurons.
abstract: Patient-specific induced pluripotent stem cells (iPSCs) show promise for use as tools for in vitro modeling of Parkinson's disease. We sought to improve the efficiency of dopaminergic (DA) neuron induction from iPSCs by the using surface markers expressed in DA progenitors to increase the significance of the phenotypic analysis. By sorting for a CD184high/CD44-fraction during neural differentiation, we obtained a population of cells that were enriched in DA neuron precursor cells and achieved higher differentiation efficiencies than those obtained through the same protocol without sorting. This high efficiency method of DA neuronal induction enabled reliable detection of reactive oxygen species (ROS) accumulation and vulnerable phenotypes in PARK2 iPSCs-derived DA neurons. We additionally established a quantitative system using the mt-mKeima reporter system to monitor mitophagy in which mitochondria fuse with lysosomes and, by combining this system with the method of DA neuronal induction described above, determined that mitophagy is impaired in PARK2 neurons. These findings suggest that the efficiency of DA neuron induction is important for the precise detection of cellular phenotypes in modeling Parkinson's disease.
authors.affiliation: Department of Physiology, Keio University, School of Medicine, Tokyo, Japan.
authors.forename: Sadafumi
authors.initials: S
authors.lastname: Suzuki
authors.affiliation: Center for Genomic and Regenerative Medicine, Juntendo University, School of Medicine, Tokyo, Japan. Electronic address: awado@juntendo.ac.jp.
authors.forename: Wado
authors.initials: W
authors.lastname: Akamatsu
authors.affiliation: Department of Physiology, Keio University, School of Medicine, Tokyo, Japan.
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Kisa
authors.affiliation: Department of Physiology, Keio University, School of Medicine, Tokyo, Japan.
authors.forename: Takefumi
authors.initials: T
authors.lastname: Sone
authors.affiliation: Center for Genomic and Regenerative Medicine, Juntendo University, School of Medicine, Tokyo, Japan; Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan.
authors.forename: Kei-Ichi
authors.initials: KI
authors.lastname: Ishikawa
authors.affiliation: Department of Pharmacology, Hoshi University, Pharmacy and Pharmaceutical Sciences, Tokyo, Japan.
authors.forename: Naoko
authors.initials: N
authors.lastname: Kuzumaki
authors.affiliation: Laboratory for Cell Function Dynamics, Brain Science Institute, RIKEN, Saitama, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Katayama
authors.affiliation: Laboratory for Cell Function Dynamics, Brain Science Institute, RIKEN, Saitama, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Miyawaki
authors.affiliation: Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan.
authors.forename: Nobutaka
authors.initials: N
authors.lastname: Hattori
authors.affiliation: Department of Physiology, Keio University, School of Medicine, Tokyo, Japan. Electronic address: hidokano@a2.keio.jp.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Okano
mesh_terms: D017209:Apoptosis; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D059290:Dopaminergic Neurons / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D063306:Mitophagy / Q000502:physiology; D008959:Models, Neurological; D058953:Neural Stem Cells / Q000166:cytology / Q000378:metabolism; D010300:Parkinson Disease / Q000209:etiology / Q000378:metabolism / Q000473:pathology; D017382:Reactive Oxygen Species / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*
chemical_list: C497483:CD44 protein, human; D018960:Hyaluronan Receptors; D017382:Reactive Oxygen Species; D044767:Ubiquitin-Protein Ligases; C111567:parkin protein
keywords: Dopaminergic neurons; Flow cytometry; Induced pluripotent stem cells; Mitophagy; Parkinson's disease