Griffin, Síle Marie (2016) Directing stem cell differentiation towards a neuronal fate using nicotinamide. Doctoral thesis, Keele University.

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Neuronal cell loss and damage in the central nervous system are characteristics of debilitating brain related-degenerative disorders such as Parkinson’s disease. The last four decades of research have focused on the promise of cell replacement therapy to replace lost cells, repair the damage and provide functional recovery in affected neural circuits. A thorough understanding of the signals implicated in the development of neurons will greatly facilitate the use of cell replacement therapy.

This project aimed to investigate the possibility of using nicotinamide, the amide form of vitamin B3, to promote the development of mature neuronal subtypes from mouse embryonic stem cells, and whether these could form a dopaminergic phenotype, to progress research in stem cell-derived therapies for Parkinson’s disease.

Treatment of mouse embryonic stem cell monolayer cultures (46C Sox1GFP reporter cell line) with nicotinamide at the early onset of development not only increased the efficiency of neuronal generation but also enriched the ratio of purified neurons to non-neuronal cells. Nicotinamide acted at the initial stages of differentiation to promote accelerated neural lineage entry by embryonic stem cells in adherent monolayer cultures. The pluripotent stem cell and neural progenitor cell populations could be reduced by treating cells with nicotinamide, which also facilitated accelerated neuronal differentiation.

Nicotinamide selectively enhanced the production of catecholaminergic, serotonergic and GABAergic neurons and, moreover, accelerated neuronal maturation. A reduction in the proportion of proliferating cells in nicotinamide-treated cultures was demonstrated– that is, nicotinamide enhanced cell-cycle exit, thereby promoting
neuronal differentiation.

The potential of nicotinamide was introduced to a novel, small-molecule-based strategy using pluripotent stem cell sources. Nicotinamide was shown to function synergistically with signalling molecules known to enhance a dopaminergic phenotype, to direct differentiating cells to adopt a dopaminergic cell fate.

Thus, novel findings suggest that nicotinamide is a key signalling factor in brain development, and is required in a definable dosage range and times for the normal
formation of dopamine neurons. This study supports previous evidence that vitamins and their metabolites play a fundamental role in neuronal development.

Item Type: Thesis (Doctoral)
Subjects: R Medicine > RC Internal medicine > RC0321 Neuroscience. Biological psychiatry. Neuropsychiatry
Divisions: Faculty of Medicine and Health Sciences > Institute for Science and Technology in Medicine
Depositing User: Lisa Bailey
Date Deposited: 31 Oct 2016 10:04
Last Modified: 31 Oct 2016 10:04

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