Abstract
This protocol describes the biofabrication of 3D millimeter-scale human muscle units, embedding non-planar muscle fibers wrapped by fibroblasts-rich endomysium and intertwined with microvascular networks. Suspended muscle fibers are formed through the self-assembly of human myoblasts within cylindrical cavities generated in a sacrificial gelatin template cast in a 3D printed frame. Following myotube differentiation, muscle fibers are embedded in a 3D matrix containing endothelial cells and muscle-derived fibroblasts. The cellular complexity of the environment is instrumental to drive fibroblast migration towards muscle fibers and to induce the organ-specific differentiation of endothelial cells. This advanced 3D muscle model can be applied to analyze the biological mechanisms underlying specific muscle diseases which involve a complex remodeling of the muscle environment (e.g., muscular dystrophies and fibrosis) whereby the pathological interplay among different cell populations drives the onset and progression of the disease.
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Acknowledgements
This project has received funding from the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement No 860715.
Images were adapted from Engineering an Environment for the Study of Fibrosis: A 3D Human Muscle Model with Endothelium Specificity and Endomysium; Bersini, Gilardi, Ugolini, Sansoni, Talò, Perego, Zanotti, Ostano, Mora, Soncini, Vanoni, Lombardi, Moretti; Cell Reports, Volume 25, 3858-3868, Copyright 2018, with permission from Elsevier, Creative Commons CC-BY-NC-ND license.
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Bersini, S., Francescato, R., Moretti, M. (2022). Biofabrication of 3D Human Muscle Model with Vascularization and Endomysium. In: Rasponi, M. (eds) Organ-on-a-Chip. Methods in Molecular Biology, vol 2373. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1693-2_13
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DOI: https://doi.org/10.1007/978-1-0716-1693-2_13
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