Modeling amyotrophic lateral sclerosis in pure human iPSc-derived motor neurons isolated by a novel FACS double selection technique
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...
| Главные авторы: | , , , , , , , , |
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| Формат: | Статья |
| Язык: | English |
| Опубликовано: |
Elsevier
2015-10-01
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| Серии: | Neurobiology of Disease |
| Предметы: | |
| Online-ссылка: | http://www.sciencedirect.com/science/article/pii/S0969996115002296 |
| _version_ | 1830187819733090304 |
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| author | Diana Toli Dorothée Buttigieg Stéphane Blanchard Thomas Lemonnier Boris Lamotte d’Incamps Sarah Bellouze Gilbert Baillat Delphine Bohl Georg Haase |
| author_facet | Diana Toli Dorothée Buttigieg Stéphane Blanchard Thomas Lemonnier Boris Lamotte d’Incamps Sarah Bellouze Gilbert Baillat Delphine Bohl Georg Haase |
| author_sort | Diana Toli |
| collection | DOAJ |
| description | 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 p75NTR surface epitope and an HB9::RFP lentivirus reporter. The p75NTR/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. |
| first_indexed | 2024-12-17T22:34:53Z |
| format | Article |
| id | doaj.art-127b0c1de7534dae9c254c3dc12a0a5d |
| institution | Directory Open Access Journal |
| issn | 1095-953X |
| language | English |
| last_indexed | 2024-12-17T22:34:53Z |
| publishDate | 2015-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Neurobiology of Disease |
| spelling | doaj.art-127b0c1de7534dae9c254c3dc12a0a5d2022-12-21T21:30:07ZengElsevierNeurobiology of Disease1095-953X2015-10-0182269280Modeling amyotrophic lateral sclerosis in pure human iPSc-derived motor neurons isolated by a novel FACS double selection techniqueDiana Toli0Dorothée Buttigieg1Stéphane Blanchard2Thomas Lemonnier3Boris Lamotte d’Incamps4Sarah Bellouze5Gilbert Baillat6Delphine Bohl7Georg Haase8Institut Pasteur and INSERM U1115, Unité Biothérapies pour les Maladies Neurodégénératives, 28 rue du Dr Roux, 75015 Paris, FranceInstitut 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, FranceInstitut Pasteur and INSERM U1115, Unité Biothérapies pour les Maladies Neurodégénératives, 28 rue du Dr Roux, 75015 Paris, FranceInstitut Pasteur and INSERM U1115, Unité Biothérapies pour les Maladies Neurodégénératives, 28 rue du Dr Roux, 75015 Paris, FranceCentre de Neurophysique, Physiologie, Pathologie, UMR 8119 CNRS and Université Paris Descartes, 45 rue des Saints-Pères, 75270 Paris Cedex 6, FranceInstitut 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, FranceInstitut 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, FranceInstitut Pasteur and INSERM U1115, Unité Biothérapies pour les Maladies Neurodégénératives, 28 rue du Dr Roux, 75015 Paris, France; Correspondence to: D. Bohl, Institut du Cerveau et de la Moelle épinière - ICM, INSERM U 1127 - CNRS UMR-7225 - UPMC-Université Paris 6 - Hôpital de la Pitié-Salpêtrière, 47, boulevard de l'Hôpital, 75013 Paris, France.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; Correspondence to: G. Haase, Institut de Neurosciences de la Timone, UMR 7289, CNRS and Aix-Marseille University, 27, boulevard Jean Moulin, 13005 Marseille cx 5, France. Fax: +33 4 9132 4056.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 p75NTR surface epitope and an HB9::RFP lentivirus reporter. The p75NTR/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.http://www.sciencedirect.com/science/article/pii/S0969996115002296Motor neuron diseaseAmyotrophic lateral sclerosis (ALS)Induced pluripotent stem cells (iPSc)Cell deathAxon degenerationFluorescent-activated cell sorting (FACS) |
| spellingShingle | Diana Toli Dorothée Buttigieg Stéphane Blanchard Thomas Lemonnier Boris Lamotte d’Incamps Sarah Bellouze Gilbert Baillat Delphine Bohl Georg Haase Modeling amyotrophic lateral sclerosis in pure human iPSc-derived motor neurons isolated by a novel FACS double selection technique Neurobiology of Disease Motor neuron disease Amyotrophic lateral sclerosis (ALS) Induced pluripotent stem cells (iPSc) Cell death Axon degeneration Fluorescent-activated cell sorting (FACS) |
| title | Modeling amyotrophic lateral sclerosis in pure human iPSc-derived motor neurons isolated by a novel FACS double selection technique |
| title_full | Modeling amyotrophic lateral sclerosis in pure human iPSc-derived motor neurons isolated by a novel FACS double selection technique |
| title_fullStr | Modeling amyotrophic lateral sclerosis in pure human iPSc-derived motor neurons isolated by a novel FACS double selection technique |
| title_full_unstemmed | Modeling amyotrophic lateral sclerosis in pure human iPSc-derived motor neurons isolated by a novel FACS double selection technique |
| title_short | Modeling amyotrophic lateral sclerosis in pure human iPSc-derived motor neurons isolated by a novel FACS double selection technique |
| title_sort | modeling amyotrophic lateral sclerosis in pure human ipsc derived motor neurons isolated by a novel facs double selection technique |
| topic | Motor neuron disease Amyotrophic lateral sclerosis (ALS) Induced pluripotent stem cells (iPSc) Cell death Axon degeneration Fluorescent-activated cell sorting (FACS) |
| url | http://www.sciencedirect.com/science/article/pii/S0969996115002296 |
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