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Decellularized Plant-Based Scaffolds for Guided Alignment of Myoblast Cells

Santiago Campuzano, Nicolette B. Mogilever, View ORCID ProfileAndrew E. Pelling
doi: https://doi.org/10.1101/2020.02.23.958686
Santiago Campuzano
‡Department of Biology, University of Ottawa, Ottawa, ON, Canada
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Nicolette B. Mogilever
‡Department of Biology, University of Ottawa, Ottawa, ON, Canada
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Andrew E. Pelling
†STEM Complex, Department of Physics, University of Ottawa, Ottawa, ON, Canada
‡Department of Biology, University of Ottawa, Ottawa, ON, Canada
§Institute for Science Society and Policy, University of Ottawa, Ottawa, ON, Canada
∥SymbioticA, School of Human Sciences, Physiology and Human Biology, University of Western Australia, Perth, WA 6009, Australia
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  • For correspondence: a@pellinglab.net
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Abstract

Alignment and orientation of cells in vivo plays a crucial role in the functionality of tissue. A challenged faced by traditional cell culture approaches is that the majority of two-dimensional substrates fail to induce a controlled alignment of cells in vitro. To address this challenge, approaches utilizing mechanical stresses, exposure to electrical fields, structurally aligned biomaterials and/or textured microfabricated substrates, have been developed to control the organization of cells through microenvironmental stimuli. In the field of muscle tissue engineering it is often desirable to control the alignment and fusion of muscle precursor cells as it more closely resembles in vivo conditions. In this study, we utilize plant-derived cellulose biomaterials to control the in vitro alignment of C2C12 murine myoblasts. We hereby report that cells display a clear sensitivity to the highly aligned vascular bundle architectures found in decellularized celery (Apium graveolens). Conveniently, the xylem and phloem channels lie within the 10-100μm diameter, which has been shown to be optimal diameter for myoblast alignment through contact guidance. Following 10 days in proliferation media, F-actin filaments were observed to be aligned parallel to the longitudinal axis of the vascular bundle. Subsequently, following 5 days in differentiation media, myoblast maintained an aligned morphology, which led to the formation of aligned myotubes. We therefore conclude that the microtopography of the vascular bundle guides muscle cell alignment. The results presented here highlight the potential of this plant-derived scaffold for in vitro studies of muscle myogenesis, where structural anisotropy is required to more closely resemble in vivo conditions.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted February 24, 2020.
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Decellularized Plant-Based Scaffolds for Guided Alignment of Myoblast Cells
Santiago Campuzano, Nicolette B. Mogilever, Andrew E. Pelling
bioRxiv 2020.02.23.958686; doi: https://doi.org/10.1101/2020.02.23.958686
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Decellularized Plant-Based Scaffolds for Guided Alignment of Myoblast Cells
Santiago Campuzano, Nicolette B. Mogilever, Andrew E. Pelling
bioRxiv 2020.02.23.958686; doi: https://doi.org/10.1101/2020.02.23.958686

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