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Optimization of an in vitro bioassay to monitor growth and formation of myotubes in real time

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dc.contributor.author Murphy, Sylvia M.
dc.contributor.author Kiely, Maeve
dc.contributor.author Jakeman, Philip M.
dc.contributor.author Kiely, Patrick A.
dc.contributor.author Carson, Brian P.
dc.contributor.other Murphy-Tighe, Sylvia
dc.date.accessioned 2016-09-02T13:48:34Z
dc.date.available 2016-09-02T13:48:34Z
dc.date.issued 2016
dc.identifier.uri http://hdl.handle.net/10344/5184
dc.description peer-reviewed en_US
dc.description.abstract The importance of growth and maintenance of skeletal muscle is vital for long term health and quality of life. Appropriate nutrition with specific bioactivities relevant to the functionalities of tissues such as skeletal muscle, can assist in maintaining and promoting adaptive responses to biological and environmental stresses which prevent muscle atrophy and promote hypertrophy. The aim of this investigation was to develop a novel in vitro cell-based electric impedance assay to study myoblast to myotube formation on the real time cell analysis (RTCA) platform (xCELLigence (TM), ACEA) and to validate the system by testing myotube responses to hypertrophic stimuli. C2C12 myoblasts were proliferated until 70% confluent in Dulbecco's Modified Eagles Medium (DMEM) (10% FBS) and subsequently differentiated to myotubes over 8 days in DMEM [2% horse serum (HS)]. Changes in cell behaviour and adhesion properties were monitored by measuring impedance via interdigitated microelectrodes in the base of E-16 cell culture dishes. To establish the suitability of this assay to monitor nutrient regulation of muscle hypertrophy, leucine, a known potent regulator of MPS was then supplemented to the fully formed myotubes in physiologically relevant conditions - 0.20 mM, 0.40 mM, 0.6 mM, 0.8 mM and above 1.0 mM, 1.5 mM, 2.0 mM and impedance subsequently monitored. Parallel experiments highlighting alterations in myotube thickness, muscle protein synthesis (MPS) (mammalian target of rapamycin; mTOR) and differentiation (myogenin) were conducted to support RTCA bioassay findings. This in vitro bioassay can be used to monitor skeletal muscle behaviour and identify nutrient compounds with bioactivities promoting skeletal muscle hypertrophy, reducing muscle atrophy and thus inform the development of novel nutrient formulations for the maintenance of skeletal muscle. en_US
dc.language.iso eng en_US
dc.publisher Portland Press en_US
dc.relation.ispartofseries Bioscience Reports;36, (3), pii: e00330
dc.rights First published by Portland Press http://dx.doi.org/10.1042/BSR20160036 en_US
dc.subject hypertrophy en_US
dc.subject MPB en_US
dc.subject MPS en_US
dc.subject RTCA en_US
dc.subject skeletal muscle en_US
dc.subject C2C12 en_US
dc.title Optimization of an in vitro bioassay to monitor growth and formation of myotubes in real time en_US
dc.type info:eu-repo/semantics/article en_US
dc.type.supercollection all_ul_research en_US
dc.type.supercollection ul_published_reviewed en_US
dc.date.updated 2016-09-02T13:39:02Z
dc.description.version PUBLISHED
dc.identifier.doi 10.1042/BSR20160036
dc.contributor.sponsor EI en_US
dc.contributor.sponsor SFI en_US
dc.relation.projectid TC20130001 en_US
dc.relation.projectid 13/CDA/2228 en_US
dc.rights.accessrights info:eu-repo/semantics/openAccess en_US
dc.internal.rssid 1645048
dc.internal.copyrightchecked Yes
dc.description.status peer-reviewed


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