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
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
Affiliations
Sonia Sanz Muñoz
Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia
Martin Engel
Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia
Rachelle Balez
Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia
Dzung Do-Ha
Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia
Mauricio Castro Cabral-da-Silva
Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia
Damian Hernández
Department of Anatomy & Neuroscience, University of Melbourne, Parkville, VIC 3010, Australia
Tracey Berg
Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia
Jennifer A. Fifita
Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, NSW 2190, Australia
Natalie Grima
Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, NSW 2190, Australia
Shu Yang
Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, NSW 2190, Australia
Ian P. Blair
Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, NSW 2190, Australia
Garth Nicholson
Concord Repatriation General Hospital, University of Sydney ANZAC Research Institute, Sydney, NSW 2006, Australia
Anthony L. Cook
Wicking Dementia Research and Education Centre, University of Tasmania, Hobart, TAS 7001, Australia
Alex W. Hewitt
Department of Surgery, University of Melbourne, Parkville, VIC 3010, Australia
Alice Pébay
Department of Anatomy & Neuroscience, University of Melbourne, Parkville, VIC 3010, Australia
Lezanne Ooi
Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia
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.
