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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MDPI AG
2022-07-01
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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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language | English |
last_indexed | 2024-03-09T10:22:14Z |
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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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