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A. Laabich, G.P. Vissvesvaran, K.L. Lieu, K. Murata, T.E. McGinn, C.C. Manmoto, J.R. Sinclair, I. Karliga, R. Kubota, A. Fawzi; Protective Effect of Crocin Against Blue and White Light Mediated Photoreceptor Cell Death in Bovine and Monkey Retinal Primary Cell Culture . Invest. Ophthalmol. Vis. Sci. 2006;47(13):4801.
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Light exposure has been attributed to the incidence of age related macular degeneration (AMD). The present study was performed to examine the effect of Crocin on light–induced rods and cones cell death in primary retinal cell cultures.
Primary retinal cell cultures were prepared from adult Monkey and Bovine retinas. Fifteen day–old cultures were exposed to blue–actinic light (900 lux, 420 nm; 1.1 mW/cm2) or to white fluorescent light (6000 lux, 2.3 mW/cm2) for 24 hours. Cultures were treated by addition of different concentrations of Crocin (10–160 µM) for 24 hours before light exposure. Cultures kept in the dark were used as controls. Sytox Green assays was used to evaluate cell death. Rods, cones and bipolar cells were immunolabeled with specific antibodies and were counted using automated imaging microscope and image–based cell count software. TUNEL labeling method was used to detect fragmented DNA in fixed cells after light exposure.
Primary retinal cell cultures prepared contained a mixture of retinal cells enriched in photoreceptors, bipolar cells and müller cells. Twenty four hours exposure to blue–actinic and white light induced 70–80% photoreceptor cell death in Bovine and Monkey retinal cell cultures. No cell death was detected in bipolar and müller cells following the light exposure. Crocin protected the photoreceptors against blue and white light mediated damage in a concentration–dependent manner with an EC50 of about 30 µM. TUNEL labeling assays confirmed light damage of the photoreceptors.
These results show that blue and white fluorescent light selectively induce rods and cones cell death in this in vitro model. Crocin protects retinal photoreceptors against blue and white light–induced cell death.
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