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Development of strategies for isolation and functional testing of cell lines for neuronal regeneration

Neves Dos Reis, Joana; (2019) Development of strategies for isolation and functional testing of cell lines for neuronal regeneration. Doctoral thesis (Ph.D), UCL (University College London). Green open access

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Abstract

In the Central Nervous System there is limited neuronal regeneration ability after injury. Olfactory Ensheathing Cells are olfactory system cells guiding neurons and supporting regeneration from the olfactory mucosa to the bulb. They also promote structural and functional recovery in vivo in animal models of CNS injury. Nevertheless, patient sample variability results in inconsistent results in autologous transplants. A universal cell therapy seems to be the envisaged solution. This project aimed to optimise the culture of OECs from rat olfactory mucosa (investigating the impact of timing of addition of NT-3 and AraC exposure for 24h) and develop in vitro assays for testing a human conditionally immortalised OEC line (PA5) for neuronal regeneration ability. Significantly higher proportion of Thy1.1-positive cells was seen when NT3 was added later (day 6 versus day 1; p<0.018). There was no significant difference in cell yield and proportion with exposure to AraC. Rat OECs were tested in co-culture with NG108 neuronal cell line. OECs generally performed similarly to the positive control F7/SC in all measurements. The same assay with PA5 cells yielded inconclusive results. Rat DRG neurons were cultured on PA5 cells and had longer neurites comparing to culture on PLL (P=0.010). Finally PA5 cells were preliminarily tested for neuronal alignment. DRG neurons co-cultured on PA5 showed a low angles-skewed distribution and a difference in the distribution of DRG angles when compared to culture on F7/SC suggesting a tendency for PA5 cells to self-align and guide neurons in in vitro. Overall this work reveals a tendency of PA5 cell line to promote regeneration of CNS neurons. Nevertheless much more work need to be undertaken to further confirm these results either using different models in vitro or in vivo or accessing other neuronal regeneration abilities.

Type: Thesis (Doctoral)
Qualification: Ph.D
Title: Development of strategies for isolation and functional testing of cell lines for neuronal regeneration
Event: UCL
Open access status: An open access version is available from UCL Discovery
Language: English
Additional information: Copyright © The Author 2019. Original content in this thesis is licensed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) Licence (https://creativecommons.org/licenses/by/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms.
UCL classification: UCL
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/10077513
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