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E Eckert, KA Kobuch, L Aigner, D Spiegel; Preservation Of Ganglion Cells In Adult Retinal Tissue Cultures - An In Vitro- Model For Neuroprotection . Invest. Ophthalmol. Vis. Sci. 2002;43(13):757.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To evaluate the potential of an in vitro-model of adult full thickness retina in perfusion culture for the examination of neuroprotection. Methods: Whole mounts of adult retina-RPE-choroid-specimens including the optic nerve head were prepared from porcine eyes within two hours after death of the animals. The tissue was transfered into a two-compartment culture container (Minucells Minutissue®) and constantly perfused with fresh medium from two sides for 4, 7 or 9 days, total number n=28. The preservation of the axonal structure was assessed histologically in flat mounts by marcation with neurofilament-antibody (NF 200, Sigma®), and by transport-activity for fluorogold (Fluka / Sigma®), which had been fixed in a gelatin sponge (Gelita, Braun®) upon the optic nerve head. Results: Under these culture conditions, the retina remained well preserved for up to 9 days. Fragmentation and destruction of the axons started in the center of the specimens at the optic nerve head after 4 days. But in the periphery, the axonal structures remained preserved for up to 9 days. Axonal transport as evidenced by transport of fluorogold from the optic nerve stump towards the ganglion cell body was still active after 4 days. Conclusion: The flat specimens of a defined area of the retina around the optic nerve allowed an assessment of nerve fiber density for a limited time period of up to 9 days. The model may thus serve as a highly differentiated and relevant in vitro system for the examination of pharmacotoxicological questions of the neural retina, especially with the option to use human tissue from corneal donor eyes.
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