Urciuolo, A;
Serena, E;
Ghua, R;
Zatti, S;
Giomo, M;
Mattei, N;
Vetralla, M;
... Elvassore, N; + view all
(2020)
Y Engineering a 3D in vitro model of human skeletal muscle at the single fiber scale.
PLOS ONE
, 15
(5)
, Article e0232081. 10.1371/journal.pone.0232081.
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Abstract
The reproduction of reliable in vitro models of human skeletal muscle is made harder by the intrinsic 3D structural complexity of this tissue. Here we coupled engineered hydrogel with 3D structural cues and specific mechanical properties to derive human 3D muscle constructs (“myobundles”) at the scale of single fibers, by using primary myoblasts or myoblasts derived from embryonic stem cells. To this aim, cell culture was performed in confined, laminin-coated micrometric channels obtained inside a 3D hydrogel characterized by the optimal stiffness for skeletal muscle myogenesis. Primary myoblasts cultured in our 3D culture system were able to undergo myotube differentiation and maturation, as demonstrated by the proper expression and localization of key components of the sarcomere and sarcolemma. Such approach allowed the generation of human myobundles of ~10 mm in length and ~120 μm in diameter, showing spontaneous contraction 7 days after cell seeding. Transcriptome analyses showed higher similarity between 3D myobundles and skeletal signature, compared to that found between 2D myotubes and skeletal muscle, mainly resulting from expression in 3D myobundles of categories of genes involved in skeletal muscle maturation, including extracellular matrix organization. Moreover, imaging analyses confirmed that structured 3D culture system was conducive to differentiation/maturation also when using myoblasts derived from embryonic stem cells. In conclusion, our structured 3D model is a promising tool for modelling human skeletal muscle in healthy and diseases conditions.
| Type: | Article |
|---|---|
| Title: | Y Engineering a 3D in vitro model of human skeletal muscle at the single fiber scale |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1371/journal.pone.0232081 |
| Publisher version: | https://doi.org/10.1371/journal.pone.0232081 |
| Language: | English |
| Additional information: | © 2020 Urciuolo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
| Keywords: | Science & Technology, Multidisciplinary Sciences, Science & Technology - Other Topics, MYOGENESIS, INJURY, REPAIR, BIOMATERIALS, HYDROGELS, LAMININ, VIVO |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health |
| URI: | https://discovery-pp.ucl.ac.uk/id/eprint/10142010 |
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