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
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 Alzhe...
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MDPI AG
2020-09-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/9/9/2018 |
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doaj-89e7ab23e46047a6a0f8fe91d1fcfe642020-11-25T03:33:07ZengMDPI AGCells2073-44092020-09-0192018201810.3390/cells9092018A Simple Differentiation Protocol for Generation of Induced Pluripotent Stem Cell-Derived Basal Forebrain-Like Cholinergic Neurons for Alzheimer’s Disease and Frontotemporal Dementia Disease ModelingSonia Sanz Muñoz0Martin Engel1Rachelle Balez2Dzung Do-Ha3Mauricio Castro Cabral-da-Silva4Damian Hernández5Tracey Berg6Jennifer A. Fifita7Natalie Grima8Shu Yang9Ian P. Blair10Garth Nicholson11Anthony L. Cook12Alex W. Hewitt13Alice Pébay14Lezanne Ooi15Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, AustraliaIllawarra Health and Medical Research Institute, Wollongong, NSW 2522, AustraliaIllawarra Health and Medical Research Institute, Wollongong, NSW 2522, AustraliaIllawarra Health and Medical Research Institute, Wollongong, NSW 2522, AustraliaIllawarra Health and Medical Research Institute, Wollongong, NSW 2522, AustraliaDepartment of Anatomy & Neuroscience, University of Melbourne, Parkville, VIC 3010, AustraliaIllawarra Health and Medical Research Institute, Wollongong, NSW 2522, AustraliaCentre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, NSW 2190, AustraliaCentre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, NSW 2190, AustraliaCentre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, NSW 2190, AustraliaCentre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, NSW 2190, AustraliaConcord Repatriation General Hospital, University of Sydney ANZAC Research Institute, Sydney, NSW 2006, AustraliaWicking Dementia Research and Education Centre, University of Tasmania, Hobart, TAS 7001, AustraliaDepartment of Surgery, University of Melbourne, Parkville, VIC 3010, AustraliaDepartment of Anatomy & Neuroscience, University of Melbourne, Parkville, VIC 3010, AustraliaIllawarra Health and Medical Research Institute, Wollongong, NSW 2522, AustraliaThe 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.https://www.mdpi.com/2073-4409/9/9/2018induced pluripotent stem cellsdisease modellingneuronal differentiationcholinergic neuronsAlzheimer’s diseasefrontotemporal dementia |
| collection |
DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
Sonia Sanz Muñoz Martin Engel Rachelle Balez Dzung Do-Ha Mauricio Castro Cabral-da-Silva Damian Hernández Tracey Berg Jennifer A. Fifita Natalie Grima Shu Yang Ian P. Blair Garth Nicholson Anthony L. Cook Alex W. Hewitt Alice Pébay Lezanne Ooi |
| spellingShingle |
Sonia Sanz Muñoz Martin Engel Rachelle Balez Dzung Do-Ha Mauricio Castro Cabral-da-Silva Damian Hernández Tracey Berg Jennifer A. Fifita Natalie Grima Shu Yang Ian P. Blair Garth Nicholson Anthony L. Cook Alex W. Hewitt Alice Pébay Lezanne Ooi 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 induced pluripotent stem cells disease modelling neuronal differentiation cholinergic neurons Alzheimer’s disease frontotemporal dementia |
| author_facet |
Sonia Sanz Muñoz Martin Engel Rachelle Balez Dzung Do-Ha Mauricio Castro Cabral-da-Silva Damian Hernández Tracey Berg Jennifer A. Fifita Natalie Grima Shu Yang Ian P. Blair Garth Nicholson Anthony L. Cook Alex W. Hewitt Alice Pébay Lezanne Ooi |
| author_sort |
Sonia Sanz Muñoz |
| title |
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 |
| title_short |
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 |
| title_full |
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 |
| title_fullStr |
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 |
| title_full_unstemmed |
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 |
| title_sort |
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 |
| publisher |
MDPI AG |
| series |
Cells |
| issn |
2073-4409 |
| publishDate |
2020-09-01 |
| description |
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. |
| topic |
induced pluripotent stem cells disease modelling neuronal differentiation cholinergic neurons Alzheimer’s disease frontotemporal dementia |
| url |
https://www.mdpi.com/2073-4409/9/9/2018 |
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