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Functional neural differentiation of adult hippocampus derived stem cells

Mazzantini, M.; (2010) Functional neural differentiation of adult hippocampus derived stem cells. Masters thesis , UCL (University College London). Green open access

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Abstract

Stem cells are defined by their capacity to differentiate into different phenotypes while maintaining their ability to replicate indefinitely. These characteristics depend on their intrinsic capabilities and their interaction with the cellular niche. However, stem cells isolated later in development show a restricted differentiation capability limited to the cell lineage specifically present in the tissue of origin. Using an adult, hippocampal-derived stem cell line (designated the CHIP stem cell line), I started looking at their integration once they had been implanted into hippocampal organotypic slices. This would have given me an idea of the sequential steps they follow during their integration and how this relates to what is known to happen in vivo. Knowing that in the hippocampus, they originate from the sub granular zone and develop into granule cells, I carried out an electrophysiological characterization of dentate gyrus granule cells both in organotypic and in acute slice. It became clear that the survival of stem cells after the implantation was affected by the culture medium used for the organotypic slices. Switching to a different culture medium improved the survival of the stem cells but at the same time they did not show signs of integration (e.g. dendritic-like protrusions). I therefore started looking at stem cell functional differentiation in vitro without the slice. In the CHIP stem cell line plus, in addition, in another hippocampal stem cell line (HCN95) I compared the response to two differentiation protocols. I employed several standard neuronal markers (including NeuN) in order to define their developmental status. Unlike most other studies I also used a marker for the alpha subunit of the voltage-activated sodium channel (Nav1.2) which is responsible for the generation of action potentials in mature neurons. Checking stem cell ability to fire action potentials at various time intervals, in parallel to the expression of neuronal markers, I found that Nav1.2 expression better correlates with the functional maturation of our neuronal stem cell lines when compared to NeuN. Therefore, Nav1.2 expression can be employed as reliable marker of mature neuronal phenotype.

Type: Thesis (Masters)
Title: Functional neural differentiation of adult hippocampus derived stem cells
Open access status: An open access version is available from UCL Discovery
Language: English
UCL classification: UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Neuro, Physiology and Pharmacology
URI: https://discovery.ucl.ac.uk/id/eprint/772653
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