Cargando…

Engineering skeletal muscle tissues with advanced maturity improves synapse formation with human induced pluripotent stem cell-derived motor neurons

To develop effective cures for neuromuscular diseases, human-relevant in vitro models of neuromuscular tissues are critically needed to probe disease mechanisms on a cellular and molecular level. However, previous attempts to co-culture motor neurons and skeletal muscle have resulted in relatively i...

Descripción completa

Detalles Bibliográficos
Autores principales: Santoso, Jeffrey W., Li, Xiling, Gupta, Divya, Suh, Gio C., Hendricks, Eric, Lin, Shaoyu, Perry, Sarah, Ichida, Justin K., Dickman, Dion, McCain, Megan L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: AIP Publishing LLC 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8282350/
https://www.ncbi.nlm.nih.gov/pubmed/34286174
http://dx.doi.org/10.1063/5.0054984
_version_ 1783722993210359808
author Santoso, Jeffrey W.
Li, Xiling
Gupta, Divya
Suh, Gio C.
Hendricks, Eric
Lin, Shaoyu
Perry, Sarah
Ichida, Justin K.
Dickman, Dion
McCain, Megan L.
author_facet Santoso, Jeffrey W.
Li, Xiling
Gupta, Divya
Suh, Gio C.
Hendricks, Eric
Lin, Shaoyu
Perry, Sarah
Ichida, Justin K.
Dickman, Dion
McCain, Megan L.
author_sort Santoso, Jeffrey W.
collection PubMed
description To develop effective cures for neuromuscular diseases, human-relevant in vitro models of neuromuscular tissues are critically needed to probe disease mechanisms on a cellular and molecular level. However, previous attempts to co-culture motor neurons and skeletal muscle have resulted in relatively immature neuromuscular junctions (NMJs). In this study, NMJs formed by human induced pluripotent stem cell (hiPSC)-derived motor neurons were improved by optimizing the maturity of the co-cultured muscle tissue. First, muscle tissues engineered from the C2C12 mouse myoblast cell line, cryopreserved primary human myoblasts, and freshly isolated primary chick myoblasts on micromolded gelatin hydrogels were compared. After three weeks, only chick muscle tissues remained stably adhered to hydrogels and exhibited progressive increases in myogenic index and stress generation, approaching values generated by native muscle tissue. After three weeks of co-culture with hiPSC-derived motor neurons, engineered chick muscle tissues formed NMJs with increasing co-localization of pre- and postsynaptic markers as well as increased frequency and magnitude of synaptic activity, surpassing structural and functional maturity of previous in vitro models. Engineered chick muscle tissues also demonstrated increased expression of genes related to sarcomere maturation and innervation over time, revealing new insights into the molecular pathways that likely contribute to enhanced NMJ formation. These approaches for engineering advanced neuromuscular tissues with relatively mature NMJs and interrogating their structure and function have many applications in neuromuscular disease modeling and drug development.
format Online
Article
Text
id pubmed-8282350
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher AIP Publishing LLC
record_format MEDLINE/PubMed
spelling pubmed-82823502021-07-19 Engineering skeletal muscle tissues with advanced maturity improves synapse formation with human induced pluripotent stem cell-derived motor neurons Santoso, Jeffrey W. Li, Xiling Gupta, Divya Suh, Gio C. Hendricks, Eric Lin, Shaoyu Perry, Sarah Ichida, Justin K. Dickman, Dion McCain, Megan L. APL Bioeng Articles To develop effective cures for neuromuscular diseases, human-relevant in vitro models of neuromuscular tissues are critically needed to probe disease mechanisms on a cellular and molecular level. However, previous attempts to co-culture motor neurons and skeletal muscle have resulted in relatively immature neuromuscular junctions (NMJs). In this study, NMJs formed by human induced pluripotent stem cell (hiPSC)-derived motor neurons were improved by optimizing the maturity of the co-cultured muscle tissue. First, muscle tissues engineered from the C2C12 mouse myoblast cell line, cryopreserved primary human myoblasts, and freshly isolated primary chick myoblasts on micromolded gelatin hydrogels were compared. After three weeks, only chick muscle tissues remained stably adhered to hydrogels and exhibited progressive increases in myogenic index and stress generation, approaching values generated by native muscle tissue. After three weeks of co-culture with hiPSC-derived motor neurons, engineered chick muscle tissues formed NMJs with increasing co-localization of pre- and postsynaptic markers as well as increased frequency and magnitude of synaptic activity, surpassing structural and functional maturity of previous in vitro models. Engineered chick muscle tissues also demonstrated increased expression of genes related to sarcomere maturation and innervation over time, revealing new insights into the molecular pathways that likely contribute to enhanced NMJ formation. These approaches for engineering advanced neuromuscular tissues with relatively mature NMJs and interrogating their structure and function have many applications in neuromuscular disease modeling and drug development. AIP Publishing LLC 2021-07-13 /pmc/articles/PMC8282350/ /pubmed/34286174 http://dx.doi.org/10.1063/5.0054984 Text en © 2021 Author(s). https://creativecommons.org/licenses/by/4.0/All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle Articles
Santoso, Jeffrey W.
Li, Xiling
Gupta, Divya
Suh, Gio C.
Hendricks, Eric
Lin, Shaoyu
Perry, Sarah
Ichida, Justin K.
Dickman, Dion
McCain, Megan L.
Engineering skeletal muscle tissues with advanced maturity improves synapse formation with human induced pluripotent stem cell-derived motor neurons
title Engineering skeletal muscle tissues with advanced maturity improves synapse formation with human induced pluripotent stem cell-derived motor neurons
title_full Engineering skeletal muscle tissues with advanced maturity improves synapse formation with human induced pluripotent stem cell-derived motor neurons
title_fullStr Engineering skeletal muscle tissues with advanced maturity improves synapse formation with human induced pluripotent stem cell-derived motor neurons
title_full_unstemmed Engineering skeletal muscle tissues with advanced maturity improves synapse formation with human induced pluripotent stem cell-derived motor neurons
title_short Engineering skeletal muscle tissues with advanced maturity improves synapse formation with human induced pluripotent stem cell-derived motor neurons
title_sort engineering skeletal muscle tissues with advanced maturity improves synapse formation with human induced pluripotent stem cell-derived motor neurons
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8282350/
https://www.ncbi.nlm.nih.gov/pubmed/34286174
http://dx.doi.org/10.1063/5.0054984
work_keys_str_mv AT santosojeffreyw engineeringskeletalmuscletissueswithadvancedmaturityimprovessynapseformationwithhumaninducedpluripotentstemcellderivedmotorneurons
AT lixiling engineeringskeletalmuscletissueswithadvancedmaturityimprovessynapseformationwithhumaninducedpluripotentstemcellderivedmotorneurons
AT guptadivya engineeringskeletalmuscletissueswithadvancedmaturityimprovessynapseformationwithhumaninducedpluripotentstemcellderivedmotorneurons
AT suhgioc engineeringskeletalmuscletissueswithadvancedmaturityimprovessynapseformationwithhumaninducedpluripotentstemcellderivedmotorneurons
AT hendrickseric engineeringskeletalmuscletissueswithadvancedmaturityimprovessynapseformationwithhumaninducedpluripotentstemcellderivedmotorneurons
AT linshaoyu engineeringskeletalmuscletissueswithadvancedmaturityimprovessynapseformationwithhumaninducedpluripotentstemcellderivedmotorneurons
AT perrysarah engineeringskeletalmuscletissueswithadvancedmaturityimprovessynapseformationwithhumaninducedpluripotentstemcellderivedmotorneurons
AT ichidajustink engineeringskeletalmuscletissueswithadvancedmaturityimprovessynapseformationwithhumaninducedpluripotentstemcellderivedmotorneurons
AT dickmandion engineeringskeletalmuscletissueswithadvancedmaturityimprovessynapseformationwithhumaninducedpluripotentstemcellderivedmotorneurons
AT mccainmeganl engineeringskeletalmuscletissueswithadvancedmaturityimprovessynapseformationwithhumaninducedpluripotentstemcellderivedmotorneurons