Myocyte Culture with Decellularized Skeletal Muscle Sheet with Observable Interaction with the Extracellular Matrix

In skeletal muscles, muscle fibers are highly organized and bundled within the basement membrane. Several microfabricated substrate models have failed to mimic the macrostructure of native muscle, including various extracellular matrix (ECM) proteins. Therefore, we developed and evaluated a system u...

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Main Authors: Satoshi Nakada, Yuri Yamashita, Seiya Akiba, Takeru Shima, Eri Arikawa-Hirasawa
Format: Article
Language:English
Published: MDPI AG 2022-07-01
Series:Bioengineering
Subjects:
Online Access:https://www.mdpi.com/2306-5354/9/7/309
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author Satoshi Nakada
Yuri Yamashita
Seiya Akiba
Takeru Shima
Eri Arikawa-Hirasawa
author_facet Satoshi Nakada
Yuri Yamashita
Seiya Akiba
Takeru Shima
Eri Arikawa-Hirasawa
author_sort Satoshi Nakada
collection DOAJ
description In skeletal muscles, muscle fibers are highly organized and bundled within the basement membrane. Several microfabricated substrate models have failed to mimic the macrostructure of native muscle, including various extracellular matrix (ECM) proteins. Therefore, we developed and evaluated a system using decellularized muscle tissue and mouse myoblasts C2C12 to analyze the interaction between native ECM and myocytes. Chicken skeletal muscle was sliced into sheets and decellularized to prepare decellularized skeletal muscle sheets (DSMS). C2C12 was then seeded and differentiated on DSMS. Immunostaining for ECM molecules was performed to examine the relationship between myoblast adhesion status, myotube orientation, and collagen IV orientation. Myotube survival in long-term culture was confirmed by calcein staining. C2C12 myoblasts adhered to scaffolds in DSMS and developed adhesion plaques and filopodia. Furthermore, C2C12 myotubes showed orientation along the ECM orientation within DSMS. Compared to plastic dishes, detachment was less likely to occur on DSMS, and long-term incubation was possible. This culture technique reproduces a cell culture environment reflecting the properties of living skeletal muscle, thereby allowing studies on the interaction between the ECM and myocytes.
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spelling doaj.art-678c63aea31f4d99b42d6dd403e9deae2023-12-01T21:53:55ZengMDPI AGBioengineering2306-53542022-07-019730910.3390/bioengineering9070309Myocyte Culture with Decellularized Skeletal Muscle Sheet with Observable Interaction with the Extracellular MatrixSatoshi Nakada0Yuri Yamashita1Seiya Akiba2Takeru Shima3Eri Arikawa-Hirasawa4Japanese Center for Research on Women in Sport, Juntendo University Graduate School of Health and Sports Science, Chiba 270-1695, JapanResearch Institute for Diseases of Old Age, Juntendo University Graduate School of Medicine, Tokyo 113-8421, JapanResearch Institute for Diseases of Old Age, Juntendo University Graduate School of Medicine, Tokyo 113-8421, JapanResearch Institute for Diseases of Old Age, Juntendo University Graduate School of Medicine, Tokyo 113-8421, JapanJapanese Center for Research on Women in Sport, Juntendo University Graduate School of Health and Sports Science, Chiba 270-1695, JapanIn skeletal muscles, muscle fibers are highly organized and bundled within the basement membrane. Several microfabricated substrate models have failed to mimic the macrostructure of native muscle, including various extracellular matrix (ECM) proteins. Therefore, we developed and evaluated a system using decellularized muscle tissue and mouse myoblasts C2C12 to analyze the interaction between native ECM and myocytes. Chicken skeletal muscle was sliced into sheets and decellularized to prepare decellularized skeletal muscle sheets (DSMS). C2C12 was then seeded and differentiated on DSMS. Immunostaining for ECM molecules was performed to examine the relationship between myoblast adhesion status, myotube orientation, and collagen IV orientation. Myotube survival in long-term culture was confirmed by calcein staining. C2C12 myoblasts adhered to scaffolds in DSMS and developed adhesion plaques and filopodia. Furthermore, C2C12 myotubes showed orientation along the ECM orientation within DSMS. Compared to plastic dishes, detachment was less likely to occur on DSMS, and long-term incubation was possible. This culture technique reproduces a cell culture environment reflecting the properties of living skeletal muscle, thereby allowing studies on the interaction between the ECM and myocytes.https://www.mdpi.com/2306-5354/9/7/309skeletal musclecell culturemyoblastsextracellular matrixdecellularizationmicrodevice
spellingShingle Satoshi Nakada
Yuri Yamashita
Seiya Akiba
Takeru Shima
Eri Arikawa-Hirasawa
Myocyte Culture with Decellularized Skeletal Muscle Sheet with Observable Interaction with the Extracellular Matrix
Bioengineering
skeletal muscle
cell culture
myoblasts
extracellular matrix
decellularization
microdevice
title Myocyte Culture with Decellularized Skeletal Muscle Sheet with Observable Interaction with the Extracellular Matrix
title_full Myocyte Culture with Decellularized Skeletal Muscle Sheet with Observable Interaction with the Extracellular Matrix
title_fullStr Myocyte Culture with Decellularized Skeletal Muscle Sheet with Observable Interaction with the Extracellular Matrix
title_full_unstemmed Myocyte Culture with Decellularized Skeletal Muscle Sheet with Observable Interaction with the Extracellular Matrix
title_short Myocyte Culture with Decellularized Skeletal Muscle Sheet with Observable Interaction with the Extracellular Matrix
title_sort myocyte culture with decellularized skeletal muscle sheet with observable interaction with the extracellular matrix
topic skeletal muscle
cell culture
myoblasts
extracellular matrix
decellularization
microdevice
url https://www.mdpi.com/2306-5354/9/7/309
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