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Murine muscle engineered from dermal precursors: an in vitro model for skeletal muscle generation, degeneration and fatty infiltration.

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Murine muscle engineered from dermal precursors: an in vitro model for skeletal muscle generation, degeneration and fatty infiltration.

dc.contributor.author	García-Parra, P.
dc.contributor.author	Naldaiz-Gastesi, N.
dc.contributor.author	Maroto, M.
dc.contributor.author	Padín, J.F.
dc.contributor.author	Goicoechea, M.
dc.contributor.author	Aiastui, A.
dc.contributor.author	Fernández-Morales, J.C.
dc.contributor.author	García Belda, Paula
dc.contributor.author	Lacalle, J.
dc.contributor.author	Alava, J.I.
dc.contributor.author	García Verdugo, José Manuel
dc.contributor.author	García, A.G.
dc.contributor.author	Izeta, A.
dc.contributor.author	López de Munain, A.
dc.date.accessioned	2014-02-19T09:24:26Z
dc.date.available	2014-12-20T09:24:26Z
dc.date.issued	2013
dc.identifier.uri	http://hdl.handle.net/10550/33057
dc.description.abstract	Skeletal muscle can be engineered by converting dermal precursors into muscle progenitors and differentiated myocytes. However, the efficiency of muscle development remains relatively low and it is currently unclear if this is due to poor characterization of the myogenic precursors, the protocols used for cell differentiation, or a combination of both. In this study, we characterized myogenic precursors present in murine dermospheres, and evaluated mature myotubes grown in a novel three-dimensional culture system. After 5<br>7 days of differentiation, we observed isolated, twitching myotubes followed by spontaneous contractions of the entire tissue-engineered muscle construct on an extracellular matrix (ECM). In vitro engineered myofibers expressed canonical muscle markers and exhibited a skeletal (not cardiac) muscle ultrastructure, with numerous striations and the presence of aligned, enlarged mitochondria, intertwined with sarcoplasmic reticula (SR). Engineered myofibers exhibited Na+- and Ca2+-dependent inward currents upon acetylcholine (ACh) stimulation and tetrodotoxin-sensitive spontaneous action potentials. Moreover, ACh, nicotine, and caffeine elicited cytosolic Ca2+ transients; fiber contractions coupled to these Ca2+ transients suggest that Ca2+ entry is activating calcium-induced calcium release from the SR. Blockade by d-tubocurarine of ACh-elicited inward currents and Ca2+ transients suggests nicotinic receptor involvement. Interestingly, after 1 month, engineered muscle constructs showed progressive degradation of the myofibers concomitant with fatty infiltration, paralleling the natural course of muscular degeneration. We conclude that mature myofibers may be differentiated on the ECM from myogenic precursor cells present in murine dermospheres, in an in vitro system that mimics some characteristics found in aging and muscular degeneration.
dc.relation.ispartof	Tissue Engineering. Part C Methods, 2013, vol. 20, num. 1, p. 28-41
dc.rights.uri	info:eu-repo/semantics/openAccess
dc.source	García-Parra, P. Naldaiz-Gastesi, N. Maroto, M. Padín, J.F. Goicoechea, M. Aiastui, A. Fernández-Morales, J.C. García Belda, Paula Lacalle, J. Alava, J.I. García Verdugo, José Manuel García, A.G. Izeta, A. López de Munain, A. 2013 Murine muscle engineered from dermal precursors: an in vitro model for skeletal muscle generation, degeneration and fatty infiltration. Tissue Engineering. Part C Methods 20 1 28 41
dc.subject	Enginyeria biomèdica
dc.subject	Biologia experimental
dc.title	Murine muscle engineered from dermal precursors: an in vitro model for skeletal muscle generation, degeneration and fatty infiltration.
dc.type	info:eu-repo/semantics/article
dc.date.updated	2014-02-19T09:24:26Z
dc.identifier.idgrec	085213