The study of retina regeneration in animal models has been of interest due to the implication it has in developing potential treatments for retinal diseases such as macular degeneration. Newts are excellent models to study regeneration because they retain the ability to regenerate their retina into adulthood. The process by which newts regenerate their retina is called retinal pigment epithelium (RPE) reprogramming. During this process, the layer of cells on the back of the eye, the RPE, will lose its identity and reprogram into retinal cells. However, the exact mechanism of how RPE undergoes reprogramming has yet to be elucidated. The purpose of this project, therefore, was to establish a cell culture (ex-vivo) system to study the factors and pathways that may be involved in the process of RPE reprogramming. The ex-vivo system was established by creating newt retina-less eye cups (RLECs) in which the anterior portion of the eye, along with the retina were removed, leaving behind the RPE. The posterior eye cup, containing RPE, was attached to a filter membrane and media containing different combinations of growth factors or small molecules that were added to induce reprogramming. The cultured RLECs were collected at 2, 4, 6 and 10 weeks and analyzed using histology, immunohistochemistry, and a cell death assay. RLECs treated with growth factors and small molecules were able to regenerate a retina-like structure that tested positive for early retinal progenitors after 10 weeks in culture, making RLECs a good and viable ex-vivo system to study RPE reprogramming.
Author: Bailey Haynes
Advisor: Dr. Katia Del Rio-Tsonis, Department of Biology