Aim: I aimed to investigate methods of delivering neuroprotection to the retina: a) using magnetofection (a non-viral transfection technique) in ocular tissue, b) using stem cells as a vector of neuroprotection delivery and c) harnessing galvanotaxis to direct migration of stem cells into the retina. Methods: For the recipient test bed of the experiments in my thesis, I used retinal explants from mice. The viability of retinal explants was determined using retinal ganglion cell health as a read-out in the form of Sholl plots. I also explored magnetofection with oscillation as a non-viral technique for neuroprotection delivery in ocular tissue using cornea and retinal explants. Using the galvanotaxis technique, I explored its use in directing neural stem cells in-vitro with an electrotactic chamber and ex-vivo on a retinal explant in a modified Boyden chamber. Finally, I magnetofected neural stem cells to over-express BDNF and directed their migration into the retina using galvanotaxis. Results: The retinal explants had a viability of up to 3 days based on the Sholl plots of retinal ganglion cells. Magnetofection with oscillation transfected cornea endothelium and the retinal ganglion cell layer with GFP in the explant models. It also transfected neural stem cells with BDNF-myc. Directed migration with neural stem cells occurred in the electrotactic chamber as well as in the retinal explant model. In the absence of electric field, no migration into the retina occurred. Neurospheres transfected with BDNF-myc also migrated into the retina when exposed to an electric field. Conclusion: Within the period of up to 3 days, retinal explants can be used to investigate neuroprotective therapeutic agents using Sholl plots of retinal ganglion cells. Magnetofection with oscillation is a novel non-viral technique for potentially transfecting the eye in the anterior and posterior segments. Neurospheres can be directed to migrate into the retina using an electric field in the ex-vivo model.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:716063 |
| Date | January 2017 |
| Creators | Ng, Wai Siene |
| Publisher | Cardiff University |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://orca.cf.ac.uk/101044/ |
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