Our society has an ageing population, and this could lead to a large economic burden if not addressed. One potential method of solving age-related medical issues is the use of mesenchymal stem cells (MSCs). The differentiation capabilities and immunomodulatory characteristics of MSCs make them an ideal target candidate for the development of stem cell therapies. So far, there has been limited success in the clinic with MSC treatments, and electrical stimulation (ES) is being investigated to better translate MSC research into effective patient outcomes. ES can alter cell migration, proliferation and differentiation, and improve wound healing; however, the mechanisms behind such effects is poorly understood. The myriad parameters involved in an ES system renders the optimisation of a successful ES regime an arduous task for researchers. In this work, the Y201 immortalised MSC line was used. Firstly, PPIA and YWHAZ were identified as a suitable pair of reference genes under ES using quantitative real-time PCR. These were verified using the "delta delta CT" method, BestKeeper, and NormFinder. Secondly, we observed a reduction in DNA quantity and no change to osteogenic differentiation with ES by day 28 in MSCs in the presence and absence of catalase, an enzyme specific to hydrogen peroxide (H2O2). This suggested that extracellular H2O2 produced at the cathode does not play a role on the long-term effects of ES, despite a reduction in the expression of antioxidant genes SRXN1 and HMOX1 in the catalase-treated group. Finally, through RNA sequencing it was found that the transcriptomic response to 3 and 6 daily ES sessions correlated highly, with a large number of genes expressed relating to not only inflammation, but also the other stages of wound healing. GPR68, a cell surface receptor, was upregulated across 1, 3, and 6 sessions of ES, suggesting its involvement in ES signal transduction. These findings could allow for more rapid optimisation of ES regimes, and thereby invoke a more widespread adoption of this technique for controlling cell behaviour.
Date of Award | 31 Dec 2023 |
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Original language | English |
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Awarding Institution | - The University of Manchester
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Supervisor | Enrique Amaya (Supervisor) & Sarah Cartmell (Supervisor) |
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Understanding the Mechanisms of Direct Current Electrical Stimulation in Human Mesenchymal Stem Cells
Steel, L. (Author). 31 Dec 2023
Student thesis: Phd