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© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.

Abstract

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 >92% motor neural progenitors and >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 4–6 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.

Details

Title
Robust Generation of Ready-to-Use Cryopreserved Motor Neurons from Human Pluripotent Stem Cells for Disease Modeling
Author
Hsiao-Chien Ting 1 ; Hong-Lin, Su 2 ; Mei-Fang, Chen 3 ; Harn, Horng-Jyh 4 ; Shinn-Zong, Lin 5   VIAFID ORCID Logo  ; Tzyy-Wen Chiou 6 ; Chia-Yu, Chang 7   VIAFID ORCID Logo 

 Bioinnovation Center, Buddhist Tzu Chi Medical Foundation, Hualien 970, Taiwan 
 Department of Life Sciences, National Chung Hsing University, Taichung 402, Taiwan 
 Department of Medical Research, Hualien Tzu Chi Hospital, Hualien 970, Taiwan 
 Bioinnovation Center, Buddhist Tzu Chi Medical Foundation, Hualien 970, Taiwan; Department of Pathology, Hualien Tzu Chi Hospital and Tzu Chi University, Hualien 970, Taiwan 
 Bioinnovation Center, Buddhist Tzu Chi Medical Foundation, Hualien 970, Taiwan; Department of Neurosurgery, Hualien Tzu Chi Hospital, Hualien 970, Taiwan 
 Department of Life Science and Graduate Institute of Biotechnology, National Dong Hwa University, Hualien 974, Taiwan 
 Bioinnovation Center, Buddhist Tzu Chi Medical Foundation, Hualien 970, Taiwan; Department of Medical Research, Hualien Tzu Chi Hospital, Hualien 970, Taiwan; Neuroscience Center, Hualien Tzu Chi Hospital, Hualien 970, Taiwan; Department of Nursing, Tzu Chi University of Science and Technology, Hualien 970, Taiwan 
First page
13462
Publication year
2022
Publication date
2022
Publisher
MDPI AG
ISSN
16616596
e-ISSN
14220067
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
2734636573
Copyright
© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.