Culturing the Retinal Pigment Epithelium
Class
Article
Department
Biological and Irrigation Engineering
Faculty Mentor
Elizabeth Vargis
Presentation Type
Oral Presentation
Abstract
Age-related nonexudative macular degeneration is the leading cause of vision loss in adults living in developed countries. Most ophthalmological research funding goes to searching for a cure for this common retinal disorder. One possible method for treating the disease, as well as testing new treatments, is the cultivation of artificially-grown retinal pigment epithelium (RPE) in vitro. The retinal pigment epithelium is an important tissue layer in the vertebrate eye and plays a large part in the development of dry (nonexudative) AMD. While culturing this cell type has been performed by researchers since the 1970s, an idealized protocol for best mimicking the conditions and morphology of the living RPE has not yet been found. Various protocols have been discovered to promote varying effects on cell viability, pigmentation, morphology, and behavior; analysis and experimentation with these procedures can lead to new discoveries and breakthroughs. In my research, I cultured RPE cells harvested from the eyeballs of pigs using a protocol modified from Singh et al (2001). I studied how a difference in tissue freshness, growth substrate, and extraction method resulted in vastly different results than those found by previous researchers, with loss of pigmentation and fibroblast-like fusiform morphologies in the cells. The modified protocol also resulted in an extension of cell life, with passaging continuing at least three sessions further than the senescence point found by Singh et al. This suggests the spontaneous production of immortalized cell lines without inducement, a phenomenon reported in other RPE culture studies. Experimentation with variation in substrate texture resulted in no changes to cell morphology or attachment rate, while cell division continued up until cryopreservation. In total, my research demonstrates the highly variable effects of environment on RPE cell development, and confirms discoveries and observations made by other researchers in the field of retinal pigment epithelial cell culture.
Start Date
4-9-2015 9:00 AM
Culturing the Retinal Pigment Epithelium
Age-related nonexudative macular degeneration is the leading cause of vision loss in adults living in developed countries. Most ophthalmological research funding goes to searching for a cure for this common retinal disorder. One possible method for treating the disease, as well as testing new treatments, is the cultivation of artificially-grown retinal pigment epithelium (RPE) in vitro. The retinal pigment epithelium is an important tissue layer in the vertebrate eye and plays a large part in the development of dry (nonexudative) AMD. While culturing this cell type has been performed by researchers since the 1970s, an idealized protocol for best mimicking the conditions and morphology of the living RPE has not yet been found. Various protocols have been discovered to promote varying effects on cell viability, pigmentation, morphology, and behavior; analysis and experimentation with these procedures can lead to new discoveries and breakthroughs. In my research, I cultured RPE cells harvested from the eyeballs of pigs using a protocol modified from Singh et al (2001). I studied how a difference in tissue freshness, growth substrate, and extraction method resulted in vastly different results than those found by previous researchers, with loss of pigmentation and fibroblast-like fusiform morphologies in the cells. The modified protocol also resulted in an extension of cell life, with passaging continuing at least three sessions further than the senescence point found by Singh et al. This suggests the spontaneous production of immortalized cell lines without inducement, a phenomenon reported in other RPE culture studies. Experimentation with variation in substrate texture resulted in no changes to cell morphology or attachment rate, while cell division continued up until cryopreservation. In total, my research demonstrates the highly variable effects of environment on RPE cell development, and confirms discoveries and observations made by other researchers in the field of retinal pigment epithelial cell culture.