title: Generation and Neuronal Differentiation of Patient-Specific Induced Pluripotent Stem Cells Derived from Niemann-Pick Type C1 Fibroblasts.
abstract: Patient-specific induced pluripotent stem cells (iPSCs) are discussed to provide a powerful tool to investigate pathological mechanisms of diseases. Moreover, such cells might be a future platform for individualized personal treatment of diseases with a broad spectrum of mutations and thus resulting in phenotypical specificities.Here, we present a protocol for the induction of induced pluripotent stem cells from patient fibroblasts with Niemann-Pick type C1 disease (NPC1). The induction is based on a retroviral system, using the "classical" transcription factors, which were described by Takahashi and colleagues in 2007. To obtain a neuronal in vitro model system of NPC1, human iPSCs were differentiated to neural progenitor cells (NPCs) and subsequently to cells of the neural lineage, namely, neurons and glial cells. iPSCs, NPCs, and terminal neuronal differentiated cells (NDCs) were characterized by means of immunocytochemistry as well as patch clamp recordings and calcium imaging to prove the functional maturation.
authors.affiliation: Albrecht-Kossel-Institute for Neuroregeneration (AKos), University of Rostock, Gehlsheimer Straße 20, 18147, Rostock, Germany.
authors.forename: Michaela
authors.initials: M
authors.lastname: Trilck
authors.affiliation: Albrecht-Kossel-Institute for Neuroregeneration (AKos), University of Rostock, Gehlsheimer Straße 20, 18147, Rostock, Germany.
authors.forename: Rayk
authors.initials: R
authors.lastname: Hübner
authors.affiliation: Albrecht-Kossel-Institute for Neuroregeneration (AKos), University of Rostock, Gehlsheimer Straße 20, 18147, Rostock, Germany. moritz.frech@med.uni-rostock.de.
authors.forename: Moritz J
authors.initials: MJ
authors.lastname: Frech
mesh_terms: D000577:Amides / Q000494:pharmacology; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D065150:Cellular Reprogramming; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D061252:Feeder Cells / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D051379:Mice; D058953:Neural Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D009457:Neuroglia / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D052556:Niemann-Pick Disease, Type C / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D018408:Patch-Clamp Techniques; D061251:Primary Cell Culture; D011725:Pyridines / Q000494:pharmacology; D012190:Retroviridae / Q000235:genetics; D012867:Skin / Q000166:cytology / Q000378:metabolism; D013724:Teratoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology
chemical_list: D000577:Amides; D015415:Biomarkers; D004791:Enzyme Inhibitors; D036341:Intercellular Signaling Peptides and Proteins; D011725:Pyridines; C108830:Y 27632
keywords: Calcium imaging; Embryoid bodies; Immunocytochemistry; Patch clamp; Patient-specific iPSCs; Pluripotency; Retroviral reprogramming; Teratoma