Wang, Weixin;
(2020)
Molecular and functional characterisation of Müller glia derived from retinal organoids formed by human pluripotent stem cells (hPSCs).
Doctoral thesis (Ph.D), UCL (University College London).
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Abstract
Previous studies have shown that transplantation of human Müller glia into animal models of retinal degeneration improves visual functions, possibly due to their neuroprotective ability. Since sourcing Müller glia from adult donor retinae is not acceptable for therapeutic use, differentiating Müller glia from human pluripotent stem cells (hPSCs) constitutes an attractive approach for developing cell-based therapies to treat retinal degenerative diseases. However, it is important to examine the molecular and functional characteristics of Müller glia derived from retinal organoids and compare these with cells isolated from adult retina before they can be considered for development of cell therapies. The transcriptomic profile of Müller glia isolated from retinal organoids formed by hPSCs was also examined and compared to that of the well-characterised adult Müller cell line MIO-M1. In addition, the effect of cytokines on the neuroprotective response of these cells, as well as the neuroprotective functionality of culture supernatants on rat retinal ganglion cells (RGCs) were assessed. RNA-sequencing revealed that hPSC-derived cells exhibited Müller glia-like transcriptomic profiles as compared to MIO-M1 and expressed a broad spectrum of genes known to confer neuroprotection. Short-term culture with TNF-α elicited a pleiotropic transcriptomic response in MIO-M1, with significant downregulation of gliotic genes and upregulation of antioxidant enzymes and immune mediators. Similarly, TNF-α significantly increased expression and release of several antioxidant enzymes in hPSC-derived Müller glia-like cells. Treatment of both, hPSC-derived or MIO-M1 cells with TGF-β1 did not change the expression of antioxidative enzymes in these cells in vitro. Culture supernatants of hPSC-derived cells induced neuroprotection of rat RGCs subjected to glutamate excitotoxicity, as shown by their better survival and structural integrity when compared to controls cultured without neurotrophic factors. Improved RGC survival was supported by the presence of various neuroprotective factors in the culture supernatants. In conclusion, the present study demonstrates that hPSC-derived cells displayed Müller glia-like characteristics and neuroprotective potential, which has laid a solid foundation to accelerate the clinical translation of Müller glia cell therapies.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | Molecular and functional characterisation of Müller glia derived from retinal organoids formed by human pluripotent stem cells (hPSCs) |
| Event: | Awarding institution: |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | Copyright © The Author 2020. 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. Access may initially be restricted at the author’s request. |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > Institute of Ophthalmology |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10116199 |
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