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    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

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    Author
    Munoz, SS; Engel, M; Balez, R; Do-Ha, D; Cabral-da-Silva, MC; Hernandez, D; Berg, T; Fifita, JA; Grima, N; Yang, S; ...
    Date
    2020-09-01
    Source Title
    Cells
    Publisher
    MDPI
    University of Melbourne Author/s
    Pebay, Alice; Hernandez De Santiago, Hector; Hewitt, Alex
    Affiliation
    Surgery (RMH)
    Centre for Eye Research Australia (CERA)
    Anatomy and Neuroscience
    Metadata
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    Document Type
    Journal Article
    Citations
    Munoz, S. S., Engel, M., Balez, R., Do-Ha, D., Cabral-da-Silva, M. C., Hernandez, D., Berg, T., Fifita, J. A., Grima, N., Yang, S., Blair, I. P., Nicholson, G., Cook, A. L., Hewitt, A. W., Pebay, A. & Ooi, L. (2020). 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. CELLS, 9 (9), https://doi.org/10.3390/cells9092018.
    Access Status
    Open Access
    URI
    http://hdl.handle.net/11343/251588
    DOI
    10.3390/cells9092018
    Abstract
    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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