期刊
CELLS
卷 9, 期 9, 页码 -出版社
MDPI
DOI: 10.3390/cells9092018
关键词
induced pluripotent stem cells; disease modelling; neuronal differentiation; cholinergic neurons; Alzheimer's disease; frontotemporal dementia
类别
资金
- National Health and Medical Research Council (NHMRC) of Australia [APP1071250, APP1095215]
- NHMRC Boosting Dementia Research Leadership Fellowship [APP1135720]
- NHMRC Senior Research Fellowship [1154389]
- National Health and Medical Research Council of Australia [1154389] Funding Source: NHMRC
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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