title: Modeling amyotrophic lateral sclerosis in pure human iPSc-derived motor neurons isolated by a novel FACS double selection technique.
abstract: Amyotrophic lateral sclerosis (ALS) is a severe and incurable neurodegenerative disease. Human motor neurons generated from induced pluripotent stem cells (iPSc) offer new perspectives for disease modeling and drug testing in ALS. In standard iPSc-derived cultures, however, the two major phenotypic alterations of ALS--degeneration of motor neuron cell bodies and axons--are often obscured by cell body clustering, extensive axon criss-crossing and presence of unwanted cell types. Here, we succeeded in isolating 100% pure and standardized human motor neurons by a novel FACS double selection based on a p75(NTR) surface epitope and an HB9::RFP lentivirus reporter. The p75(NTR)/HB9::RFP motor neurons survive and grow well without forming clusters or entangled axons, are electrically excitable, contain ALS-relevant motor neuron subtypes and form functional connections with co-cultured myotubes. Importantly, they undergo rapid and massive cell death and axon degeneration in response to mutant SOD1 astrocytes. These data demonstrate the potential of FACS-isolated human iPSc-derived motor neurons for improved disease modeling and drug testing in ALS and related motor neuron diseases.
authors.affiliation: Institut Pasteur and INSERM U1115, Unité Biothérapies pour les Maladies Neurodégénératives, 28 rue du Dr Roux, 75015 Paris, France.
authors.forename: Diana
authors.initials: D
authors.lastname: Toli
authors.affiliation: Institut de Neurosciences de la Timone, UMR 7289 CNRS and Aix-Marseille Université, Modélisation et Thérapie des Maladies Dégénératives des Motoneurones, 27 boulevard Jean Moulin, 13005 Marseille Cedex 5, France.
authors.forename: Dorothée
authors.initials: D
authors.lastname: Buttigieg
authors.affiliation: Institut Pasteur and INSERM U1115, Unité Biothérapies pour les Maladies Neurodégénératives, 28 rue du Dr Roux, 75015 Paris, France.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Blanchard
authors.affiliation: Institut Pasteur and INSERM U1115, Unité Biothérapies pour les Maladies Neurodégénératives, 28 rue du Dr Roux, 75015 Paris, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Lemonnier
authors.affiliation: Centre de Neurophysique, Physiologie, Pathologie, UMR 8119 CNRS and Université Paris Descartes, 45 rue des Saints-Pères, 75270 Paris Cedex 6, France.
authors.forename: Boris
authors.initials: B
authors.lastname: Lamotte d'Incamps
authors.affiliation: Institut de Neurosciences de la Timone, UMR 7289 CNRS and Aix-Marseille Université, Modélisation et Thérapie des Maladies Dégénératives des Motoneurones, 27 boulevard Jean Moulin, 13005 Marseille Cedex 5, France.
authors.forename: Sarah
authors.initials: S
authors.lastname: Bellouze
authors.affiliation: Institut de Neurosciences de la Timone, UMR 7289 CNRS and Aix-Marseille Université, Modélisation et Thérapie des Maladies Dégénératives des Motoneurones, 27 boulevard Jean Moulin, 13005 Marseille Cedex 5, France.
authors.forename: Gilbert
authors.initials: G
authors.lastname: Baillat
authors.affiliation: Institut Pasteur and INSERM U1115, Unité Biothérapies pour les Maladies Neurodégénératives, 28 rue du Dr Roux, 75015 Paris, France. Electronic address: delphine.bohl@icm-institute.org.
authors.forename: Delphine
authors.initials: D
authors.lastname: Bohl
authors.affiliation: Institut de Neurosciences de la Timone, UMR 7289 CNRS and Aix-Marseille Université, Modélisation et Thérapie des Maladies Dégénératives des Motoneurones, 27 boulevard Jean Moulin, 13005 Marseille Cedex 5, France. Electronic address: georg.haase@univ-amu.fr.
authors.forename: Georg
authors.initials: G
authors.lastname: Haase
mesh_terms: D000328:Adult; D000690:Amyotrophic Lateral Sclerosis / Q000473:pathology / Q000503:physiopathology; D001253:Astrocytes / Q000473:pathology / Q000502:physiology; D001369:Axons / Q000473:pathology / Q000502:physiology; D002470:Cell Survival; D002478:Cells, Cultured; D002648:Child; D018920:Coculture Techniques; D005434:Flow Cytometry / Q000379:methods*; D017930:Genes, Reporter; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000502:physiology; D016086:Lentivirus; D009046:Motor Neurons / Q000473:pathology / Q000502:physiology; D009154:Mutation; D009410:Nerve Degeneration / Q000473:pathology / Q000503:physiopathology; D009419:Nerve Tissue Proteins / Q000378:metabolism; D017475:Receptors, Nerve Growth Factor / Q000378:metabolism; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism; D000072105:Superoxide Dismutase-1
chemical_list: C484065:NGFR protein, human; D009419:Nerve Tissue Proteins; D017475:Receptors, Nerve Growth Factor; C000606290:SOD1 protein, human; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1
keywords: Amyotrophic lateral sclerosis (ALS); Axon degeneration; Cell death; Fluorescent-activated cell sorting (FACS); HB9; Induced pluripotent stem cells (iPSc); Motor neuron disease; Superoxide dismutase 1 (SOD1); p75NTR