Enhancing the function of iPS-derived neurons: implications for disease modelling

Rushton, David 2014. Enhancing the function of iPS-derived neurons: implications for disease modelling. PhD Thesis, Cardiff University. Item availability restricted.

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Abstract

A critical deficit in many studies using iPS cell-derived neurons is the electrophysiological properties of these cells. Using short differentiation protocols this deficit is more significant, likely due to a lack of differentiated astrocytes in these cultures. As a strategy for combatting this deficit, this study utilises astrocyte secreted factors, in the form of astrocyte conditioned medium (ACM). ACM generated cultures of neurons producing spontaneous activity more reminiscent of neurons in vivo after only 3 weeks of differentiation. Further, this study finds that voltage activated Ca2+ currents are enhanced at a very early time point using ACM and these channels, along with GABAA receptors, are vital to neuronal functional maturation. This demonstrates an endogenous mechanism present in the early stages of functional development, where neurons exhibit excitatory responses to GABA which drive the activation of voltage activated Ca2+ channels. In addition to finding that astrocyte secreted factors evoke a gain of this endogenous, activity led mechanism for functional maturation, this study also investigates methods of enhance functional maturation by manipulating Ca2+ influx. However, in the absence of ACM, direct manipulation of this endogenous mechanism appeared limited by GABA becoming an inhibitory neurotransmitter as the cells functional matured. These strategies for enhance functional maturation are then assessed using iPS cell-lines generated for a Huntington disease study, finding that GABAA and increased Ca2+ concentration in the medium evoked both increased and more consistent functional properties in theses neurons at week 2. In addition to developing enhanced protocols for neuronal differentiation, a novel protocol for producing cultures of iPS cell-derived astrocytes in the absence neurons. These cultures could provide an invaluable tool, alongside iPS cell-derived neurons, for modelling neurodegenerative disease mechanisms in both cell types.

Item Type:	Thesis (PhD)
Status:	Unpublished
Schools:	Biosciences
Subjects:	Q Science > QP Physiology
Uncontrolled Keywords:	iPS cells, GABA, astrocytes, neuronal differentiation, modelling, Huntington's, astrocyte differentiation
Funders:	Medical Research Council UK, Cure Huntington's Disease Initiative USA
Date of First Compliant Deposit:	30 March 2016
Last Modified:	10 Jan 2024 09:48
URI:	https://orca.cardiff.ac.uk/id/eprint/61275

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