title: Highly Expandable Human iPS Cell-Derived Neural Progenitor Cells (NPC) and Neurons for Central Nervous System Disease Modeling and High-Throughput Screening.
abstract: Reprogramming of human somatic cells into induced pluripotent stem (iPS) cells has greatly expanded the set of research tools available to investigate the molecular and cellular mechanisms underlying central nervous system (CNS) disorders. Realizing the promise of iPS cell technology for the identification of novel therapeutic targets and for high-throughput drug screening requires implementation of methods for the large-scale production of defined CNS cell types. Here we describe a protocol for generating stable, highly expandable, iPS cell-derived CNS neural progenitor cells (NPC) using multi-dimensional fluorescence activated cell sorting (FACS) to purify NPC defined by cell surface markers. In addition, we describe a rapid, efficient, and reproducible method for generating excitatory cortical-like neurons from these NPC through inducible expression of the pro-neural transcription factor Neurogenin 2 (iNgn2-NPC). Finally, we describe methodology for the use of iNgn2-NPC for probing human neuroplasticity and mechanisms underlying CNS disorders using high-content, single-cell-level automated microscopy assays. © 2017 by John Wiley & Sons, Inc.
authors.affiliation: Chemical Neurobiology Laboratory, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.|Center for Human Genetic Research, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.|Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.|Department of Psychiatry, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.
authors.forename: Chialin
authors.initials: C
authors.lastname: Cheng
authors.affiliation: Chemical Neurobiology Laboratory, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.|Center for Human Genetic Research, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.|Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.|Department of Psychiatry, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Fass
authors.affiliation: Harvard Stem Cell Institute and Center for Regenerative Medicine Flow Cytometry Core Facility, Massachusetts General Hospital, Boston, Massachusetts.|Currently at FACS & Imaging Core Facility, Max Planck Institute for Biology of Ageing, Cologne, Germany.
authors.forename: Kat
authors.initials: K
authors.lastname: Folz-Donahue
authors.affiliation: Center for Human Genetic Research, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.|Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.
authors.forename: Marcy E
authors.initials: ME
authors.lastname: MacDonald
authors.affiliation: Chemical Neurobiology Laboratory, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.|Center for Human Genetic Research, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.|Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.|Department of Psychiatry, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Haggarty
mesh_terms: D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D002493:Central Nervous System Diseases / Q000188:drug therapy* / Q000473:pathology*; D057166:High-Throughput Screening Assays; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D008954:Models, Biological; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D009473:Neuronal Plasticity; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D059010:Single-Cell Analysis
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D009419:Nerve Tissue Proteins; C491183:NEUROG2 protein, human
keywords: drug discovery; high-content imaging; high-throughput screening; induced pluripotent stem cell; neurodegeneration; neuroplasticity; neuropsychiatric