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Methods affecting neuronal differentiation of human adult and pluripotent stem cells

Thwaites, JW; (2015) Methods affecting neuronal differentiation of human adult and pluripotent stem cells. Doctoral thesis , UCL (University College London). Green open access

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Abstract

Stem cells have significant potential to treat many age-related degenerative disorders that affect increasing numbers of people globally. This thesis investigated the capacity for omnicytes and human pluripotent stem cells (hPSC) to undergo directed differentiation towards neuronal cell types for the treatment of ischemic stroke and Parkinson’s disease respectively. Omnicytes express a range of markers related to pluripotency and plasticity; however they are a challenging cell source to use in the development of cell therapies. Variability in omnicyte quality was associated with patient source, disease type and cryopreservation, all of which affected the reproducibility of data. Successful generation of dopaminergic neurons was achieved using a suspension-based hPSC culture system, with modified culture medium designed to replicate endogenous signalling during development. Neurons expressing key markers of dopaminergic neurons were generated and were capable of producing dopamine in response to KCl challenge. The work also showed that transfection of saRNA could enhance the expression of key genes i.e. foxa2, lmx1a and TH, relative to mock transfected cultures, although not significantly. Results also showed that the specific hESC line used (Shef6) had a greater propensity for differentiation toward dopaminergic neurons than MSUH001 hiPSC. This work successfully used saRNA to enhance gene expression, but shows that transfection efficiency is a limiting factor to its use. However, if transfection efficiency can be addressed, saRNA will become a powerful tool in the generation of cell therapies, particularly if it can be applied to suspension cell cultures.

Type: Thesis (Doctoral)
Title: Methods affecting neuronal differentiation of human adult and pluripotent stem cells
Open access status: An open access version is available from UCL Discovery
Language: English
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/1467088
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