Abstract
Degeneration of the retina can lead ultimately to devastating irreversible vision loss, such as in inherited retinitis pigmentosa and age-related macular degeneration. Currently there is no cure to prevent retinal degeneration. Quantitative cell-based assays can be used to test potential drugs that prevent the death of retinal cells. Here, we describe in detail three semi-automated cell-based protocols to identify retinoprotective factors with two retinal cell lines, rat R28 cells and mouse 661W cells. In these protocols, cells are induced to undergo death by photo-oxidation stress, growth factor depletion or cytotoxicity with sodium iodate. Pigment epithelium-derived factor, an established neurotrophic factor for retinal cells, was used as a positive control. We discuss how these protocols will prove useful in high-throughput quantitative screening to identify novel therapeutics for retinal disorders.