A Simple Differentiation Protocol for Generation of Induced Pluripotent Stem Cell-Derived Basal Forebrain-Like Cholinergic Neurons for Alzheimer's Disease and Frontotemporal Dementia Disease Modeling.

Tytuł:: A Simple Differentiation Protocol for Generation of Induced Pluripotent Stem Cell-Derived Basal Forebrain-Like Cholinergic Neurons for Alzheimer's Disease and Frontotemporal Dementia Disease Modeling.
Autorzy:: Muñoz SS; Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia.; School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, NSW 2522, Australia.
Engel M; Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia.; School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, NSW 2522, Australia.
Balez R; Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia.; School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, NSW 2522, Australia.
Do-Ha D; Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia.; School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, NSW 2522, Australia.
Cabral-da-Silva MC; Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia.; School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, NSW 2522, Australia.
Hernández D; Department of Anatomy & Neuroscience, University of Melbourne, Parkville, VIC 3010, Australia.; Department of Surgery, University of Melbourne, Parkville, VIC 3010, Australia.; Centre for Eye Research Australia, Royal Victoria Eye and Ear Hospital, Melbourne, VIC 3002, Australia.
Berg T; Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia.; School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, NSW 2522, Australia.
Fifita JA; Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, NSW 2190, Australia.
Grima N; Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, NSW 2190, Australia.
Yang S; Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, NSW 2190, Australia.
Blair IP; Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, NSW 2190, Australia.
Nicholson G; Concord Repatriation General Hospital, University of Sydney ANZAC Research Institute, Sydney, NSW 2006, Australia.
Cook AL; Wicking Dementia Research and Education Centre, University of Tasmania, Hobart, TAS 7001, Australia.
Hewitt AW; Department of Surgery, University of Melbourne, Parkville, VIC 3010, Australia.; Centre for Eye Research Australia, Royal Victoria Eye and Ear Hospital, Melbourne, VIC 3002, Australia.; School of Medicine, Menzies Institute for Medical Research, University of Tasmania, Hobart, TAS 7000, Australia.
Pébay A; Department of Anatomy & Neuroscience, University of Melbourne, Parkville, VIC 3010, Australia.; Department of Surgery, University of Melbourne, Parkville, VIC 3010, Australia.; Centre for Eye Research Australia, Royal Victoria Eye and Ear Hospital, Melbourne, VIC 3002, Australia.
Ooi L; Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia.; School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, NSW 2522, Australia.
Źródło:: Cells [Cells] 2020 Sep 02; Vol. 9 (9). Date of Electronic Publication: 2020 Sep 02.
Typ publikacji:: Journal Article; Research Support, Non-U.S. Gov't
Język:: English
Imprint Name(s):: Original Publication: Basel, Switzerland : MDPI
MeSH Terms:: Cell Differentiation/*drug effects
Cholinergic Neurons/*drug effects
Culture Media/*pharmacology
Embryoid Bodies/*drug effects
Induced Pluripotent Stem Cells/*drug effects
Primary Cell Culture/*methods
Alzheimer Disease/genetics ; Alzheimer Disease/metabolism ; Alzheimer Disease/pathology ; Amyotrophic Lateral Sclerosis/genetics ; Amyotrophic Lateral Sclerosis/metabolism ; Amyotrophic Lateral Sclerosis/pathology ; Basal Forebrain/metabolism ; Basal Forebrain/pathology ; Benzamides/pharmacology ; Brain-Derived Neurotrophic Factor/pharmacology ; Cell Line ; Cholinergic Neurons/cytology ; Cholinergic Neurons/metabolism ; Culture Media/chemistry ; Dioxoles/pharmacology ; Embryoid Bodies/cytology ; Embryoid Bodies/metabolism ; Female ; Fibroblast Growth Factor 2/pharmacology ; Frontotemporal Dementia/genetics ; Frontotemporal Dementia/metabolism ; Frontotemporal Dementia/pathology ; Growth Differentiation Factor 2/pharmacology ; Hedgehog Proteins/pharmacology ; Humans ; Induced Pluripotent Stem Cells/cytology ; Induced Pluripotent Stem Cells/metabolism ; Male ; Models, Biological ; Nerve Growth Factor/pharmacology ; Patch-Clamp Techniques ; Pyrazoles/pharmacology ; Pyrimidines/pharmacology ; Transforming Growth Factor beta/pharmacology
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Contributed Indexing:: Keywords: Alzheimer’s disease; cholinergic neurons; disease modelling; frontotemporal dementia; induced pluripotent stem cells; neuronal differentiation
Substance Nomenclature:: 0 (4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide)
0 (Benzamides)
0 (Brain-Derived Neurotrophic Factor)
0 (Culture Media)
0 (Dioxoles)
0 (GDF2 protein, human)
0 (Growth Differentiation Factor 2)
0 (Hedgehog Proteins)
0 (LDN 193189)
0 (NGF protein, human)
0 (Pyrazoles)
0 (Pyrimidines)
0 (SHH protein, human)
0 (Transforming Growth Factor beta)
103107-01-3 (Fibroblast Growth Factor 2)
7171WSG8A2 (BDNF protein, human)
9061-61-4 (Nerve Growth Factor)
Entry Date(s):: Date Created: 20200905 Date Completed: 20210325 Latest Revision: 20210325
Update Code:: 20240104
PubMed Central ID:: PMC7564334
DOI:: 10.3390/cells9092018
PMID:: 32887382
: Czasopismo naukowe

Pełny tekst

The study of neurodegenerative diseases using pluripotent stem cells requires new methods to assess neurodevelopment and neurodegeneration of specific neuronal subtypes. The cholinergic system, characterized by its use of the neurotransmitter acetylcholine, is one of the first to degenerate in Alzheimer's disease and is also affected in frontotemporal dementia. We developed a differentiation protocol to generate basal forebrain-like cholinergic neurons (BFCNs) from induced pluripotent stem cells (iPSCs) aided by the use of small molecule inhibitors and growth factors. Ten iPSC lines were successfully differentiated into BFCNs using this protocol. The neuronal cultures were characterised through RNA and protein expression, and functional analysis of neurons was confirmed by whole-cell patch clamp. We have developed a reliable protocol using only small molecule inhibitors and growth factors, while avoiding transfection or cell sorting methods, to achieve a BFCN culture that expresses the characteristic markers of cholinergic neurons.

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