A Cryopreservation Strategy for Myoblast Storage in Paper‐Based Scaffolds for Inter‐Laboratory Studies of Skeletal Muscle Health

Abstract 3D tissue‐engineered models are poised to facilitate understanding of skeletal muscle pathophysiology and identify novel therapeutic agents to improve muscle health. Adopting these culture models within the broader biology community is a challenge as many models involve complex methodologie...

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Main Authors: Saifedine T. Rjaibi, Erik Jacques, Jiaru Ni, Bin Xu, Sonya Kouthouridis, Julie Sitolle, Heta Lad, Nitya Gulati, Nancy T. Li, Henry Ahn, Howard J. Ginsberg, Boyang Zhang, Fabien Le Grand, Penney M. Gilbert, Alison P. McGuigan
Format: Article
Language:English
Published: Wiley-VCH 2024-11-01
Series:Advanced Materials Interfaces
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Online Access:https://doi.org/10.1002/admi.202400382
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author Saifedine T. Rjaibi
Erik Jacques
Jiaru Ni
Bin Xu
Sonya Kouthouridis
Julie Sitolle
Heta Lad
Nitya Gulati
Nancy T. Li
Henry Ahn
Howard J. Ginsberg
Boyang Zhang
Fabien Le Grand
Penney M. Gilbert
Alison P. McGuigan
author_facet Saifedine T. Rjaibi
Erik Jacques
Jiaru Ni
Bin Xu
Sonya Kouthouridis
Julie Sitolle
Heta Lad
Nitya Gulati
Nancy T. Li
Henry Ahn
Howard J. Ginsberg
Boyang Zhang
Fabien Le Grand
Penney M. Gilbert
Alison P. McGuigan
author_sort Saifedine T. Rjaibi
collection DOAJ
description Abstract 3D tissue‐engineered models are poised to facilitate understanding of skeletal muscle pathophysiology and identify novel therapeutic agents to improve muscle health. Adopting these culture models within the broader biology community is a challenge as many models involve complex methodologies and significant investments of time and resources to optimize manufacturing protocols. To alleviate this barrier, a protocol with commercially available reagents is developed to cryopreserve myoblasts in a 96‐well compatible format that allows tissues to be transferred to users without expertise in 2D or 3D skeletal muscle cell culture. This report validates that myoblasts encapsulated in a hydrogel and cryopreserved in paper‐based scaffolds maintain cell viability, differentiation, and function via acetylcholine‐induced transient calcium responses. Furthermore, successful shipping of myoblasts cryopreserved in paper‐based scaffolds to intra‐provincial and international collaborators is demonstrated who successfully thaw, culture, and use the 3D muscle tissues. Finally, the application of this method is confirmed for studying muscle endogenous repair by seeding freshly isolated skeletal muscle stem cells to cryopreserved then differentiated and injured tissues, demonstrating expected responses to a known stimulator of muscle stem cell self‐renewal, p38α/β MAPKi. Altogether, the 3D myoblast cryopreservation protocol offers broadened access of a complex skeletal muscle tissue model to the research community.
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spelling doaj-art-af8cfeb3963e4a71b3f90e035d3dfdee2025-08-20T02:33:01ZengWiley-VCHAdvanced Materials Interfaces2196-73502024-11-011133n/an/a10.1002/admi.202400382A Cryopreservation Strategy for Myoblast Storage in Paper‐Based Scaffolds for Inter‐Laboratory Studies of Skeletal Muscle HealthSaifedine T. Rjaibi0Erik Jacques1Jiaru Ni2Bin Xu3Sonya Kouthouridis4Julie Sitolle5Heta Lad6Nitya Gulati7Nancy T. Li8Henry Ahn9Howard J. Ginsberg10Boyang Zhang11Fabien Le Grand12Penney M. Gilbert13Alison P. McGuigan14Department of Chemical Engineering and Applied Chemistry University of Toronto Toronto ON M5S3E4 CanadaDonnelly Centre for Cellular and Biomolecular Research University of Toronto Toronto ON M5S3E1 CanadaDivision of Engineering Science University of Toronto Toronto ON M5S2E4 CanadaDonnelly Centre for Cellular and Biomolecular Research University of Toronto Toronto ON M5S3E1 CanadaDepartment of Chemical Engineering McMaster University Hamilton ON L8S4L7 CanadaInstitut NeuroMyoGène Pathophysiology and Genetics of Neuron and Muscle (PGNM) Unit Université Claude Bernard Lyon 1 CNRS UMR 5261, INSERM U1315 Lyon 69008 FranceDonnelly Centre for Cellular and Biomolecular Research University of Toronto Toronto ON M5S3E1 CanadaDepartment of Chemical Engineering and Applied Chemistry University of Toronto Toronto ON M5S3E4 CanadaDepartment of Chemical Engineering and Applied Chemistry University of Toronto Toronto ON M5S3E4 CanadaDepartment of Surgery University of Toronto Toronto ON M5T 1P5 CanadaDepartment of Surgery University of Toronto Toronto ON M5T 1P5 CanadaDepartment of Chemical Engineering McMaster University Hamilton ON L8S4L7 CanadaInstitut NeuroMyoGène Pathophysiology and Genetics of Neuron and Muscle (PGNM) Unit Université Claude Bernard Lyon 1 CNRS UMR 5261, INSERM U1315 Lyon 69008 FranceDonnelly Centre for Cellular and Biomolecular Research University of Toronto Toronto ON M5S3E1 CanadaDepartment of Chemical Engineering and Applied Chemistry University of Toronto Toronto ON M5S3E4 CanadaAbstract 3D tissue‐engineered models are poised to facilitate understanding of skeletal muscle pathophysiology and identify novel therapeutic agents to improve muscle health. Adopting these culture models within the broader biology community is a challenge as many models involve complex methodologies and significant investments of time and resources to optimize manufacturing protocols. To alleviate this barrier, a protocol with commercially available reagents is developed to cryopreserve myoblasts in a 96‐well compatible format that allows tissues to be transferred to users without expertise in 2D or 3D skeletal muscle cell culture. This report validates that myoblasts encapsulated in a hydrogel and cryopreserved in paper‐based scaffolds maintain cell viability, differentiation, and function via acetylcholine‐induced transient calcium responses. Furthermore, successful shipping of myoblasts cryopreserved in paper‐based scaffolds to intra‐provincial and international collaborators is demonstrated who successfully thaw, culture, and use the 3D muscle tissues. Finally, the application of this method is confirmed for studying muscle endogenous repair by seeding freshly isolated skeletal muscle stem cells to cryopreserved then differentiated and injured tissues, demonstrating expected responses to a known stimulator of muscle stem cell self‐renewal, p38α/β MAPKi. Altogether, the 3D myoblast cryopreservation protocol offers broadened access of a complex skeletal muscle tissue model to the research community.https://doi.org/10.1002/admi.202400382cryopreservationin vitromyoblastsregenerationskeletal muscletissue engineering
spellingShingle Saifedine T. Rjaibi
Erik Jacques
Jiaru Ni
Bin Xu
Sonya Kouthouridis
Julie Sitolle
Heta Lad
Nitya Gulati
Nancy T. Li
Henry Ahn
Howard J. Ginsberg
Boyang Zhang
Fabien Le Grand
Penney M. Gilbert
Alison P. McGuigan
A Cryopreservation Strategy for Myoblast Storage in Paper‐Based Scaffolds for Inter‐Laboratory Studies of Skeletal Muscle Health
Advanced Materials Interfaces
cryopreservation
in vitro
myoblasts
regeneration
skeletal muscle
tissue engineering
title A Cryopreservation Strategy for Myoblast Storage in Paper‐Based Scaffolds for Inter‐Laboratory Studies of Skeletal Muscle Health
title_full A Cryopreservation Strategy for Myoblast Storage in Paper‐Based Scaffolds for Inter‐Laboratory Studies of Skeletal Muscle Health
title_fullStr A Cryopreservation Strategy for Myoblast Storage in Paper‐Based Scaffolds for Inter‐Laboratory Studies of Skeletal Muscle Health
title_full_unstemmed A Cryopreservation Strategy for Myoblast Storage in Paper‐Based Scaffolds for Inter‐Laboratory Studies of Skeletal Muscle Health
title_short A Cryopreservation Strategy for Myoblast Storage in Paper‐Based Scaffolds for Inter‐Laboratory Studies of Skeletal Muscle Health
title_sort cryopreservation strategy for myoblast storage in paper based scaffolds for inter laboratory studies of skeletal muscle health
topic cryopreservation
in vitro
myoblasts
regeneration
skeletal muscle
tissue engineering
url https://doi.org/10.1002/admi.202400382
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