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Validation of a Three-Dimensional Culture System for the Differentiation of Multipotential Mesenchymal Stromal Cells by Uniaxial Strain

Parker, Lindsey Christine (2017) Validation of a Three-Dimensional Culture System for the Differentiation of Multipotential Mesenchymal Stromal Cells by Uniaxial Strain. PhD thesis, University of Leeds.

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Abstract

The differentiation potential of multipotential mesenchymal stromal cells is known to be affected by many aspects of the cellular microenvironment, including soluble factors, extracellular matrix composition, the Young’s modulus of the substrate, cellular neighbours and externally applied forces. Despite this, reasonable understanding of harnessing soluble factors only exists. Few studies have investigated mechanotransduction in hMSC, and those published to date primarily employ unsuitable substrates, that do not facilitate the cellular adhesions known to be active in force transmission. In this study, porcine pericardium was decellularised for use as a biologically-relevant, threedimensional scaffold for the mechanostimulation of hMSC in a uniaxial strain bioreactor. Tissue stocks (n=67) were successfully decellularised and confirm biocompatible, sterile and free of contaminating genomic DNA. Histoarchitecture comparable to that of native tissue was also maintained. Tencell-specific seeding rings were found to release cytotoxic residue, and an alternative, nontoxic seeding approach was developed. The Tencell bioreactor was initially unable to maintain cell viability as a culture system, and was validated with respect to chamber humidity, culture temperature and arm displacement. Temperature maintenance was inadequate prior to re-engineering of the heating apparatus and was rectified through the use of an autotunable module. Losses of cell viability were still observed following validation as a result of medium pH changes. A Tencell culture regime utilising the HEPES buffer was successfully developed for the culture of hMSC. No significant differences in gene expression between strained and unstrained samples were found, and the greatest effects were observed between unseeded and other sample types. Additionally, seeded hMSC did not penetrate the scaffold. Overall, this study investigated the differentiation potential of hMSC cultured in a threedimensional scaffold. The Tencell bioreactor was fully validated for use as a uniaxial strain mechanostimulation device, and could be used in future studies to investigate the effect of different frequencies and magnitudes of cyclic strain.

Item Type: Thesis (PhD)
Keywords: Uniaxial strain, bioreactor, MSC, mechanotransduction
Academic Units: The University of Leeds > Faculty of Engineering (Leeds) > School of Mechanical Engineering (Leeds) > Institute of Medical and Biological Engineering (iMBE)(Leeds)
Identification Number/EthosID: uk.bl.ethos.729435
Depositing User: Dr Lindsey Parker
Date Deposited: 29 Nov 2017 14:05
Last Modified: 25 Jul 2018 09:56
URI: http://etheses.whiterose.ac.uk/id/eprint/18665

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