{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T15:44:31Z","timestamp":1788277471122,"version":"build-2803163510"},"reference-count":40,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2022,11,3]],"date-time":"2022-11-03T00:00:00Z","timestamp":1667433600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Hualien Tzu Chi Hospital","award":["IMAR-110-01-02"],"award-info":[{"award-number":["IMAR-110-01-02"]}]},{"name":"Hualien Tzu Chi Hospital","award":["MOST 107-2314-B-303-001-"],"award-info":[{"award-number":["MOST 107-2314-B-303-001-"]}]},{"name":"Hualien Tzu Chi Hospital","award":["MOST 108-2314-B-303-009-"],"award-info":[{"award-number":["MOST 108-2314-B-303-009-"]}]},{"name":"Hualien Tzu Chi Hospital","award":["MOST 109-2314-B-303-023-"],"award-info":[{"award-number":["MOST 109-2314-B-303-023-"]}]},{"name":"Hualien Tzu Chi Hospital","award":["MOST 110-2314-B-303-025-"],"award-info":[{"award-number":["MOST 110-2314-B-303-025-"]}]},{"name":"Hualien Tzu Chi Hospital","award":["MOST 111-2314-B-303 -027-MY2"],"award-info":[{"award-number":["MOST 111-2314-B-303 -027-MY2"]}]},{"name":"National Science and Technology Council","award":["IMAR-110-01-02"],"award-info":[{"award-number":["IMAR-110-01-02"]}]},{"name":"National Science and Technology Council","award":["MOST 107-2314-B-303-001-"],"award-info":[{"award-number":["MOST 107-2314-B-303-001-"]}]},{"name":"National Science and Technology Council","award":["MOST 108-2314-B-303-009-"],"award-info":[{"award-number":["MOST 108-2314-B-303-009-"]}]},{"name":"National Science and Technology Council","award":["MOST 109-2314-B-303-023-"],"award-info":[{"award-number":["MOST 109-2314-B-303-023-"]}]},{"name":"National Science and Technology Council","award":["MOST 110-2314-B-303-025-"],"award-info":[{"award-number":["MOST 110-2314-B-303-025-"]}]},{"name":"National Science and Technology Council","award":["MOST 111-2314-B-303 -027-MY2"],"award-info":[{"award-number":["MOST 111-2314-B-303 -027-MY2"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Human pluripotent stem cell (hPSC)-derived motor neurons (MNs) act as models for motor neuron diseases (MNDs), such as amyotrophic lateral sclerosis (ALS) or spinal muscular atrophy. However, the MN differentiation efficiency and viability following cryopreservation require further development for application in large-scale studies and drug screening. Here, we developed a robust protocol to convert hPSCs into MN cryopreservation stocks (hPSCs were converted into &gt;92% motor neural progenitors and &gt;91% MNs). Near-mature MNs were cryopreserved at a high thawing survival rate and 89% MN marker expression on day 32. Moreover, these MNs exhibited classical electrophysiological properties and neuromuscular junction (NMJ) formation ability within only 4\u20136 days after thawing. To apply this platform as an MND model, MN stocks were generated from SOD1G85R, SOD1G85G isogenic control, and sporadic ALS hPSC lines. The thawed ALS MNs expressed ALS-specific cytopathies, including SOD1 protein aggregation and TDP-43 redistribution. Thus, a stable and robust protocol was developed to generate ready-to-use cryopreserved MNs without further neuronal maturation processes for application in MND mechanistic studies, NMJ model establishment, and large-scale drug screening.<\/jats:p>","DOI":"10.3390\/ijms232113462","type":"journal-article","created":{"date-parts":[[2022,11,4]],"date-time":"2022-11-04T03:28:10Z","timestamp":1667532490000},"page":"13462","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Robust Generation of Ready-to-Use Cryopreserved Motor Neurons from Human Pluripotent Stem Cells for Disease Modeling"],"prefix":"10.3390","volume":"23","author":[{"given":"Hsiao-Chien","family":"Ting","sequence":"first","affiliation":[{"name":"Bioinnovation Center, Buddhist Tzu Chi Medical Foundation, Hualien 970, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hong-Lin","family":"Su","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, National Chung Hsing University, Taichung 402, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mei-Fang","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Medical Research, Hualien Tzu Chi Hospital, Hualien 970, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Horng-Jyh","family":"Harn","sequence":"additional","affiliation":[{"name":"Bioinnovation Center, Buddhist Tzu Chi Medical Foundation, Hualien 970, Taiwan"},{"name":"Department of Pathology, Hualien Tzu Chi Hospital and Tzu Chi University, Hualien 970, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4601-9933","authenticated-orcid":false,"given":"Shinn-Zong","family":"Lin","sequence":"additional","affiliation":[{"name":"Bioinnovation Center, Buddhist Tzu Chi Medical Foundation, Hualien 970, Taiwan"},{"name":"Department of Neurosurgery, Hualien Tzu Chi Hospital, Hualien 970, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tzyy-Wen","family":"Chiou","sequence":"additional","affiliation":[{"name":"Department of Life Science and Graduate Institute of Biotechnology, National Dong Hwa University, Hualien 974, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3585-5944","authenticated-orcid":false,"given":"Chia-Yu","family":"Chang","sequence":"additional","affiliation":[{"name":"Bioinnovation Center, Buddhist Tzu Chi Medical Foundation, Hualien 970, Taiwan"},{"name":"Department of Medical Research, Hualien Tzu Chi Hospital, Hualien 970, Taiwan"},{"name":"Neuroscience Center, Hualien Tzu Chi Hospital, Hualien 970, Taiwan"},{"name":"Department of Nursing, Tzu Chi University of Science and Technology, Hualien 970, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,11,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1038\/nrn3430","article-title":"The changing scene of amyotrophic lateral sclerosis","volume":"14","author":"Robberecht","year":"2013","journal-title":"Nat. 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