A single-step three-dimensional human skeletal muscle model for enhanced translational research

Three-dimensional (3D) skeletal muscle models have attracted considerable attention due to their ability to reflect the complexity of skeletal muscle structure and mimic a wide range of physiological processes in vitro. However, 3D cultures are usually associated with a higher experimental complexit...

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Main Authors: Sergio Perez-Diaz, Jaime Granado Leon, Susana López-Ortiz, Mirko Mandić, Mikael Altun, Håkan Rundqvist, Thomas Gustafsson, Rodrigo Fernandez-Gonzalo
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:Results in Engineering
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Online Access:http://www.sciencedirect.com/science/article/pii/S2590123025016408
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author Sergio Perez-Diaz
Jaime Granado Leon
Susana López-Ortiz
Mirko Mandić
Mikael Altun
Håkan Rundqvist
Thomas Gustafsson
Rodrigo Fernandez-Gonzalo
author_facet Sergio Perez-Diaz
Jaime Granado Leon
Susana López-Ortiz
Mirko Mandić
Mikael Altun
Håkan Rundqvist
Thomas Gustafsson
Rodrigo Fernandez-Gonzalo
author_sort Sergio Perez-Diaz
collection DOAJ
description Three-dimensional (3D) skeletal muscle models have attracted considerable attention due to their ability to reflect the complexity of skeletal muscle structure and mimic a wide range of physiological processes in vitro. However, 3D cultures are usually associated with a higher experimental complexity and cost. To overcome these limitations, we aimed to develop a serum-free 3D model of human skeletal muscle using skeletal muscle progenitor cells from human donors in a single step. After 10 days in serum-free myogenic differentiation medium, the 3D model showed greater Myosin heavy chain 1 (MyH1) expression compared to 0 and 5 days of differentiation and to classical 2D models. The 3D model responded to treatment with the atrophic agent myostatin with a reduction in MyH1 content. We also tested the regenerative capacity of the 3D model. MyH1 content decreased 0.16 days (4 h) after a damaging insult with barium chloride. In parallel, the 3D model increased the expression of cytokines supporting skeletal muscle repair, such as IL-6, and 12 days after injury it was able to increase MyH1 levels even above pre-injury levels. Furthermore, we tested and confirmed the effects of individual donors on the properties of our 3D model of skeletal muscle and showed that it is a suitable tool to study the interactions between different cell types. The model presented in this study shows clear advantages over similar 3D models, as it allows the replication of skeletal muscle biological processes in vitro without the complexity of advanced equipment or materials. The model responds to stimulation with external factors, appears to resemble the donor phenotype, can be used to study cell-to-cell interactions, and shows the expected response to biological processes such as muscle injury and hormonal treatment in vitro.
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spelling doaj-art-c0c5d7ce839b43e9ae14f271c1e65d822025-08-20T03:10:27ZengElsevierResults in Engineering2590-12302025-06-012610557010.1016/j.rineng.2025.105570A single-step three-dimensional human skeletal muscle model for enhanced translational researchSergio Perez-Diaz0Jaime Granado Leon1Susana López-Ortiz2Mirko Mandić3Mikael Altun4Håkan Rundqvist5Thomas Gustafsson6Rodrigo Fernandez-Gonzalo7Department of Laboratory Medicine, Division of Clinical Physiology, Karolinska Institutet, Stockholm, Sweden; Unit of Clinical Physiology, Karolinska University Hospital, Stockholm, Sweden; Corresponding author.Department of Laboratory Medicine, Division of Clinical Physiology, Karolinska Institutet, Stockholm, Sweden; Unit of Clinical Physiology, Karolinska University Hospital, Stockholm, SwedenDepartment of Laboratory Medicine, Division of Clinical Physiology, Karolinska Institutet, Stockholm, Sweden; i+HeALTH Strategic Research Group, Miguel de Cervantes European University, Valladolid, SpainDepartment of Laboratory Medicine, Division of Clinical Physiology, Karolinska Institutet, Stockholm, Sweden; Unit of Clinical Physiology, Karolinska University Hospital, Stockholm, SwedenDepartment of Laboratory Medicine, Division of Clinical Physiology, Karolinska Institutet, Stockholm, Sweden; Unit of Clinical Physiology, Karolinska University Hospital, Stockholm, SwedenDepartment of Laboratory Medicine, Division of Clinical Physiology, Karolinska Institutet, Stockholm, Sweden; Unit of Clinical Physiology, Karolinska University Hospital, Stockholm, SwedenDepartment of Laboratory Medicine, Division of Clinical Physiology, Karolinska Institutet, Stockholm, Sweden; Unit of Clinical Physiology, Karolinska University Hospital, Stockholm, SwedenDepartment of Laboratory Medicine, Division of Clinical Physiology, Karolinska Institutet, Stockholm, Sweden; Unit of Clinical Physiology, Karolinska University Hospital, Stockholm, SwedenThree-dimensional (3D) skeletal muscle models have attracted considerable attention due to their ability to reflect the complexity of skeletal muscle structure and mimic a wide range of physiological processes in vitro. However, 3D cultures are usually associated with a higher experimental complexity and cost. To overcome these limitations, we aimed to develop a serum-free 3D model of human skeletal muscle using skeletal muscle progenitor cells from human donors in a single step. After 10 days in serum-free myogenic differentiation medium, the 3D model showed greater Myosin heavy chain 1 (MyH1) expression compared to 0 and 5 days of differentiation and to classical 2D models. The 3D model responded to treatment with the atrophic agent myostatin with a reduction in MyH1 content. We also tested the regenerative capacity of the 3D model. MyH1 content decreased 0.16 days (4 h) after a damaging insult with barium chloride. In parallel, the 3D model increased the expression of cytokines supporting skeletal muscle repair, such as IL-6, and 12 days after injury it was able to increase MyH1 levels even above pre-injury levels. Furthermore, we tested and confirmed the effects of individual donors on the properties of our 3D model of skeletal muscle and showed that it is a suitable tool to study the interactions between different cell types. The model presented in this study shows clear advantages over similar 3D models, as it allows the replication of skeletal muscle biological processes in vitro without the complexity of advanced equipment or materials. The model responds to stimulation with external factors, appears to resemble the donor phenotype, can be used to study cell-to-cell interactions, and shows the expected response to biological processes such as muscle injury and hormonal treatment in vitro.http://www.sciencedirect.com/science/article/pii/S2590123025016408Skeletal muscleSkeletal muscle 3D modelsSkeletal muscle regenerationSkeletal muscle stem cellsTranslational researchTissue engineering
spellingShingle Sergio Perez-Diaz
Jaime Granado Leon
Susana López-Ortiz
Mirko Mandić
Mikael Altun
Håkan Rundqvist
Thomas Gustafsson
Rodrigo Fernandez-Gonzalo
A single-step three-dimensional human skeletal muscle model for enhanced translational research
Results in Engineering
Skeletal muscle
Skeletal muscle 3D models
Skeletal muscle regeneration
Skeletal muscle stem cells
Translational research
Tissue engineering
title A single-step three-dimensional human skeletal muscle model for enhanced translational research
title_full A single-step three-dimensional human skeletal muscle model for enhanced translational research
title_fullStr A single-step three-dimensional human skeletal muscle model for enhanced translational research
title_full_unstemmed A single-step three-dimensional human skeletal muscle model for enhanced translational research
title_short A single-step three-dimensional human skeletal muscle model for enhanced translational research
title_sort single step three dimensional human skeletal muscle model for enhanced translational research
topic Skeletal muscle
Skeletal muscle 3D models
Skeletal muscle regeneration
Skeletal muscle stem cells
Translational research
Tissue engineering
url http://www.sciencedirect.com/science/article/pii/S2590123025016408
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