An in vitro model for assessment of skeletal muscle adaptation following exercise related physiological cues

5.00
Hdl Handle:
http://hdl.handle.net/10547/311732
Title:
An in vitro model for assessment of skeletal muscle adaptation following exercise related physiological cues
Authors:
Player, Darren James
Abstract:
The aim of this Thesis was to further characterise and utilise an in vitro skeletal muscle (SkM) model, to investigate its potential use in further understanding the cellular and molecular adaptations to exercise in vivo. Candidate genes and proteins have been identified using in vivo, ex vivo and targeted in vitro experiments, however the complete picture of these molecular mechanisms are far from understood. Furthermore, the extent to which mechanical signals contribute to the intra-cellular mechanisms associated with exercise is also underinvesitgated. To this end, developing an in vitro model of SkM that can recapitulate in vivo SkM and respond to mechanical stimulation in a similar way to exercise will provide a means to begin to delineate the complex cellular and molecular regulation of SkM. The initial investigation (Chapter 3) characterised an optimal seeding density and culture period of C2C12 myoblasts within a 3 ml collagen gel. These data provided support for the use of collagen constructs seeded at 4 x 106 cells/ml, with no statistical differences observed in peak force, rate of force development and relative force compared to other seeding densities examined (table 3-2, all p > 0.05). However the use of 4 x 106 cells/ml supports previous data in a larger construct volume model, whilst the highest cell density possible in the system increases cell-cell contact required for fusion. Immunohistochemical and gene expression analyses provided evidence for the fusion of single seeded myoblasts into multinucleate myotubes, demonstrating an in vivo-like architecture. Chapter 4 presented data towards the characterisation and use of two distinct cyclical stretch regimens with respect to the acute biochemical and transcriptional responses. Data revealed increases in peak media lactate and reductions in peak media glucose, following cyclical stetch compared to control (p = 0.000 and p = 0.001 respectively, Fig. 4-2). Changes in mtDNA (Fig. 4-5) and associated mRNA transcriptional signals (Fig. 4-7) were mode dependent.
Citation:
Player, D.J. (2012) 'An in vitro model for assessment of skeletal muscle adaptation following exercise related physiological cues'. PhD thesis. University of Bedfordshire.
Publisher:
University of Bedfordshire
Issue Date:
Oct-2013
URI:
http://hdl.handle.net/10547/311732
Type:
Thesis or dissertation
Language:
en
Description:
A thesis submitted to the University of Bedfordshire, in partial fulfilment of the requirements for the degree of Doctor of Philosophy
Appears in Collections:
PhD e-theses; PhD e-theses

Full metadata record

DC FieldValue Language
dc.contributor.authorPlayer, Darren Jamesen
dc.date.accessioned2014-01-23T13:29:56Z-
dc.date.available2014-01-23T13:29:56Z-
dc.date.issued2013-10-
dc.identifier.citationPlayer, D.J. (2012) 'An in vitro model for assessment of skeletal muscle adaptation following exercise related physiological cues'. PhD thesis. University of Bedfordshire.en
dc.identifier.urihttp://hdl.handle.net/10547/311732-
dc.descriptionA thesis submitted to the University of Bedfordshire, in partial fulfilment of the requirements for the degree of Doctor of Philosophyen
dc.description.abstractThe aim of this Thesis was to further characterise and utilise an in vitro skeletal muscle (SkM) model, to investigate its potential use in further understanding the cellular and molecular adaptations to exercise in vivo. Candidate genes and proteins have been identified using in vivo, ex vivo and targeted in vitro experiments, however the complete picture of these molecular mechanisms are far from understood. Furthermore, the extent to which mechanical signals contribute to the intra-cellular mechanisms associated with exercise is also underinvesitgated. To this end, developing an in vitro model of SkM that can recapitulate in vivo SkM and respond to mechanical stimulation in a similar way to exercise will provide a means to begin to delineate the complex cellular and molecular regulation of SkM. The initial investigation (Chapter 3) characterised an optimal seeding density and culture period of C2C12 myoblasts within a 3 ml collagen gel. These data provided support for the use of collagen constructs seeded at 4 x 106 cells/ml, with no statistical differences observed in peak force, rate of force development and relative force compared to other seeding densities examined (table 3-2, all p > 0.05). However the use of 4 x 106 cells/ml supports previous data in a larger construct volume model, whilst the highest cell density possible in the system increases cell-cell contact required for fusion. Immunohistochemical and gene expression analyses provided evidence for the fusion of single seeded myoblasts into multinucleate myotubes, demonstrating an in vivo-like architecture. Chapter 4 presented data towards the characterisation and use of two distinct cyclical stretch regimens with respect to the acute biochemical and transcriptional responses. Data revealed increases in peak media lactate and reductions in peak media glucose, following cyclical stetch compared to control (p = 0.000 and p = 0.001 respectively, Fig. 4-2). Changes in mtDNA (Fig. 4-5) and associated mRNA transcriptional signals (Fig. 4-7) were mode dependent.en
dc.language.isoenen
dc.publisherUniversity of Bedfordshireen
dc.subjectC600 Sports Scienceen
dc.subjectskeletal muscleen
dc.subjectmuscle regenerationen
dc.titleAn in vitro model for assessment of skeletal muscle adaptation following exercise related physiological cuesen
dc.typeThesis or dissertationen
dc.type.qualificationnamePhDen_GB
dc.type.qualificationlevelPhDen
dc.publisher.institutionUniversity of Bedfordshireen
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