{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T04:14:55Z","timestamp":1788495295275,"version":"build-2803163510"},"reference-count":40,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2020,9,2]],"date-time":"2020-09-02T00:00:00Z","timestamp":1599004800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","award":["APP1135720"],"award-info":[{"award-number":["APP1135720"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","award":["APP1071250"],"award-info":[{"award-number":["APP1071250"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","award":["APP1095215"],"award-info":[{"award-number":["APP1095215"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","award":["1154389"],"award-info":[{"award-number":["1154389"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>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\u2019s 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.<\/jats:p>","DOI":"10.3390\/cells9092018","type":"journal-article","created":{"date-parts":[[2020,9,2]],"date-time":"2020-09-02T09:29:28Z","timestamp":1599038968000},"page":"2018","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":38,"title":["A Simple Differentiation Protocol for Generation of Induced Pluripotent Stem Cell-Derived Basal Forebrain-Like Cholinergic Neurons for Alzheimer\u2019s Disease and Frontotemporal Dementia Disease Modeling"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0970-3637","authenticated-orcid":false,"given":"Sonia Sanz","family":"Mu\u00f1oz","sequence":"first","affiliation":[{"name":"Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia"},{"name":"School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, NSW 2522, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7602-1340","authenticated-orcid":false,"given":"Martin","family":"Engel","sequence":"additional","affiliation":[{"name":"Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia"},{"name":"School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, NSW 2522, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2787-7107","authenticated-orcid":false,"given":"Rachelle","family":"Balez","sequence":"additional","affiliation":[{"name":"Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia"},{"name":"School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, NSW 2522, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dzung","family":"Do-Ha","sequence":"additional","affiliation":[{"name":"Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia"},{"name":"School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, NSW 2522, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3607-9158","authenticated-orcid":false,"given":"Mauricio Castro","family":"Cabral-da-Silva","sequence":"additional","affiliation":[{"name":"Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia"},{"name":"School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, NSW 2522, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Damian","family":"Hern\u00e1ndez","sequence":"additional","affiliation":[{"name":"Department of Anatomy &amp; Neuroscience, University of Melbourne, Parkville, VIC 3010, Australia"},{"name":"Department of Surgery, University of Melbourne, Parkville, VIC 3010, Australia"},{"name":"Centre for Eye Research Australia, Royal Victoria Eye and Ear Hospital, Melbourne, VIC 3002, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tracey","family":"Berg","sequence":"additional","affiliation":[{"name":"Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia"},{"name":"School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, NSW 2522, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer A.","family":"Fifita","sequence":"additional","affiliation":[{"name":"Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, NSW 2190, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6434-7706","authenticated-orcid":false,"given":"Natalie","family":"Grima","sequence":"additional","affiliation":[{"name":"Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, NSW 2190, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shu","family":"Yang","sequence":"additional","affiliation":[{"name":"Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, NSW 2190, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ian P.","family":"Blair","sequence":"additional","affiliation":[{"name":"Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, NSW 2190, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Garth","family":"Nicholson","sequence":"additional","affiliation":[{"name":"Concord Repatriation General Hospital, University of Sydney ANZAC Research Institute, Sydney, NSW 2006, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anthony L.","family":"Cook","sequence":"additional","affiliation":[{"name":"Wicking Dementia Research and Education Centre, University of Tasmania, Hobart, TAS 7001, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5123-5999","authenticated-orcid":false,"given":"Alex W.","family":"Hewitt","sequence":"additional","affiliation":[{"name":"Department of Surgery, University of Melbourne, Parkville, VIC 3010, Australia"},{"name":"Centre for Eye Research Australia, Royal Victoria Eye and Ear Hospital, Melbourne, VIC 3002, Australia"},{"name":"School of Medicine, Menzies Institute for Medical Research, University of Tasmania, Hobart, TAS 7000, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7408-9453","authenticated-orcid":false,"given":"Alice","family":"P\u00e9bay","sequence":"additional","affiliation":[{"name":"Department of Anatomy &amp; 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