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140751 - A simple differentiation protocol for generation of induced pluripotent.pdf (2.56 MB)

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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posted on 2023-05-20, 17:29 authored by Munoz, SS, Engel, M, Balez, R, Do-Ha, D, Cabral-da-Silva, MC, Hernandez, D, Berg, T, Fifita, JA, Grima, N, Yang, S, Blair, IP, Nicholson, G, Anthony CookAnthony Cook, Alexander HewittAlexander Hewitt, Pebay, A, Ooi, L
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.

History

Publication title

Cells

Volume

9

Issue

9

Article number

E2018

Number

E2018

Pagination

1-17

ISSN

2073-4409

Department/School

Wicking Dementia Research Education Centre

Publisher

M D P I AG

Place of publication

Switzerland

Rights statement

Copyright 2020 the authors. Licensed under Creative Commons Attribution 4.0 International (CC BY 4.0) https://creativecommons.org/licenses/by/4.0/

Repository Status

  • Open

Socio-economic Objectives

Clinical health not elsewhere classified