Cargando…

Optimization of Long-Term Human iPSC-Derived Spinal Motor Neuron Culture Using a Dendritic Polyglycerol Amine-Based Substrate

Human induced pluripotent stem cells (hiPSCs) derived from healthy and diseased individuals can give rise to many cell types, facilitating the study of mechanisms of development, human disease modeling, and early drug target validation. In this context, experimental model systems based on hiPSC-deri...

Descripción completa

Detalles Bibliográficos
Autores principales: Thiry, Louise, Clément, Jean-Pierre, Haag, Rainer, Kennedy, Timothy E, Stifani, Stefano
Formato: Online Artículo Texto
Lenguaje:English
Publicado: SAGE Publications 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8784909/
https://www.ncbi.nlm.nih.gov/pubmed/35023784
http://dx.doi.org/10.1177/17590914211073381
_version_ 1784638844914630656
author Thiry, Louise
Clément, Jean-Pierre
Haag, Rainer
Kennedy, Timothy E
Stifani, Stefano
author_facet Thiry, Louise
Clément, Jean-Pierre
Haag, Rainer
Kennedy, Timothy E
Stifani, Stefano
author_sort Thiry, Louise
collection PubMed
description Human induced pluripotent stem cells (hiPSCs) derived from healthy and diseased individuals can give rise to many cell types, facilitating the study of mechanisms of development, human disease modeling, and early drug target validation. In this context, experimental model systems based on hiPSC-derived motor neurons (MNs) have been used to study MN diseases such as spinal muscular atrophy and amyotrophic lateral sclerosis. Modeling MN disease using hiPSC-based approaches requires culture conditions that can recapitulate in a dish the events underlying differentiation, maturation, aging, and death of MNs. Current hiPSC-derived MN-based applications are often hampered by limitations in our ability to monitor MN morphology, survival, and other functional properties over a prolonged timeframe, underscoring the need for improved long-term culture conditions. Here we describe a cytocompatible dendritic polyglycerol amine (dPGA) substrate-based method for prolonged culture of hiPSC-derived MNs. We provide evidence that MNs cultured on dPGA-coated dishes are more amenable to long-term study of cell viability, molecular identity, and spontaneous network electrophysiological activity. The present study has the potential to improve hiPSC-based studies of human MN biology and disease. We describe the use of a new coating substrate providing improved conditions for long-term cultures of human iPSC-derived motor neurons, thus allowing evaluation of cell viability, molecular identity, spontaneous network electrophysiological activity, and single-cell RNA sequencing of mature motor neurons.
format Online
Article
Text
id pubmed-8784909
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher SAGE Publications
record_format MEDLINE/PubMed
spelling pubmed-87849092022-01-25 Optimization of Long-Term Human iPSC-Derived Spinal Motor Neuron Culture Using a Dendritic Polyglycerol Amine-Based Substrate Thiry, Louise Clément, Jean-Pierre Haag, Rainer Kennedy, Timothy E Stifani, Stefano ASN Neuro Original Papers Human induced pluripotent stem cells (hiPSCs) derived from healthy and diseased individuals can give rise to many cell types, facilitating the study of mechanisms of development, human disease modeling, and early drug target validation. In this context, experimental model systems based on hiPSC-derived motor neurons (MNs) have been used to study MN diseases such as spinal muscular atrophy and amyotrophic lateral sclerosis. Modeling MN disease using hiPSC-based approaches requires culture conditions that can recapitulate in a dish the events underlying differentiation, maturation, aging, and death of MNs. Current hiPSC-derived MN-based applications are often hampered by limitations in our ability to monitor MN morphology, survival, and other functional properties over a prolonged timeframe, underscoring the need for improved long-term culture conditions. Here we describe a cytocompatible dendritic polyglycerol amine (dPGA) substrate-based method for prolonged culture of hiPSC-derived MNs. We provide evidence that MNs cultured on dPGA-coated dishes are more amenable to long-term study of cell viability, molecular identity, and spontaneous network electrophysiological activity. The present study has the potential to improve hiPSC-based studies of human MN biology and disease. We describe the use of a new coating substrate providing improved conditions for long-term cultures of human iPSC-derived motor neurons, thus allowing evaluation of cell viability, molecular identity, spontaneous network electrophysiological activity, and single-cell RNA sequencing of mature motor neurons. SAGE Publications 2022-01-13 /pmc/articles/PMC8784909/ /pubmed/35023784 http://dx.doi.org/10.1177/17590914211073381 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by-nc/4.0/This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access page (https://us.sagepub.com/en-us/nam/open-access-at-sage).
spellingShingle Original Papers
Thiry, Louise
Clément, Jean-Pierre
Haag, Rainer
Kennedy, Timothy E
Stifani, Stefano
Optimization of Long-Term Human iPSC-Derived Spinal Motor Neuron Culture Using a Dendritic Polyglycerol Amine-Based Substrate
title Optimization of Long-Term Human iPSC-Derived Spinal Motor Neuron Culture Using a Dendritic Polyglycerol Amine-Based Substrate
title_full Optimization of Long-Term Human iPSC-Derived Spinal Motor Neuron Culture Using a Dendritic Polyglycerol Amine-Based Substrate
title_fullStr Optimization of Long-Term Human iPSC-Derived Spinal Motor Neuron Culture Using a Dendritic Polyglycerol Amine-Based Substrate
title_full_unstemmed Optimization of Long-Term Human iPSC-Derived Spinal Motor Neuron Culture Using a Dendritic Polyglycerol Amine-Based Substrate
title_short Optimization of Long-Term Human iPSC-Derived Spinal Motor Neuron Culture Using a Dendritic Polyglycerol Amine-Based Substrate
title_sort optimization of long-term human ipsc-derived spinal motor neuron culture using a dendritic polyglycerol amine-based substrate
topic Original Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8784909/
https://www.ncbi.nlm.nih.gov/pubmed/35023784
http://dx.doi.org/10.1177/17590914211073381
work_keys_str_mv AT thirylouise optimizationoflongtermhumanipscderivedspinalmotorneuroncultureusingadendriticpolyglycerolaminebasedsubstrate
AT clementjeanpierre optimizationoflongtermhumanipscderivedspinalmotorneuroncultureusingadendriticpolyglycerolaminebasedsubstrate
AT haagrainer optimizationoflongtermhumanipscderivedspinalmotorneuroncultureusingadendriticpolyglycerolaminebasedsubstrate
AT kennedytimothye optimizationoflongtermhumanipscderivedspinalmotorneuroncultureusingadendriticpolyglycerolaminebasedsubstrate
AT stifanistefano optimizationoflongtermhumanipscderivedspinalmotorneuroncultureusingadendriticpolyglycerolaminebasedsubstrate