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.

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.
Authors	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
Journal	Cells (Cells) Vol. 9 Issue 9 (09 02 2020) ISSN: 2073-4409 [Electronic] Switzerland
PMID	32887382 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References	4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide Benzamides Brain-Derived Neurotrophic Factor Culture Media Dioxoles GDF2 protein, human Growth Differentiation Factor 2 Hedgehog Proteins LDN 193189 NGF protein, human Pyrazoles Pyrimidines SHH protein, human Transforming Growth Factor beta Fibroblast Growth Factor 2 BDNF protein, human Nerve Growth Factor
Topics	Alzheimer Disease (genetics, metabolism, pathology) Amyotrophic Lateral Sclerosis (genetics, metabolism, pathology) Basal Forebrain (metabolism, pathology) Benzamides (pharmacology) Brain-Derived Neurotrophic Factor (pharmacology) Cell Differentiation (drug effects) Cell Line Cholinergic Neurons (cytology, drug effects, metabolism) Culture Media (chemistry, pharmacology) Dioxoles (pharmacology) Embryoid Bodies (cytology, drug effects, metabolism) Female Fibroblast Growth Factor 2 (pharmacology) Frontotemporal Dementia (genetics, metabolism, pathology) Growth Differentiation Factor 2 (pharmacology) Hedgehog Proteins (pharmacology) Humans Induced Pluripotent Stem Cells (cytology, drug effects, metabolism) Male Models, Biological Nerve Growth Factor (pharmacology) Patch-Clamp Techniques Primary Cell Culture (methods) Pyrazoles (pharmacology) Pyrimidines (pharmacology) Transforming Growth Factor beta (pharmacology)

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