Programm                 "Degeneration und Regeneration– Grundlagen, Diagnostik und Therapie"

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An Organotypic Retinal Culture as a Tool for Preclinical Drug Testing

Kohler K., Arango-Gonzales B., Pinzon-Duarte G., Guenther E.
Experimental Ophthalmology, University Eye Hospital Tübingen

Purpose: Preclinical drug tests in animals are costly and time consuming whereas conventional cell culture systems fail to reproduce many important features of an intact tissue, in particular when complex neuronal interactions have to be considered, e.g. in hereditary and age related retinal degeneration, diabetic retinopathy, and glaucoma. To overcome these limitations, we developed an organotypic culture system of the vascularized mammalian retina in which the influence of drugs could be studied in vitro under conditions in which the cellular interaction and three-dimensionality of the tissue was preserved during the whole culturing period.
Method: Under sterile conditions the eyes from neonatal rats were enucleated and the retinas dissected free, with or without the adhering RPE. The entire tissue was transferred onto the nylon membrane of a transwell chamber and maintained at the medium/gas interface to keep it wet without covering it. The cultures were placed without shaking or rotation in an incubator with adjusted atmosphere, humidity, and temperature. The medium was changed in regular intervals.
Results: Retinas were kept in the organ culture system for up to three weeks, offering the possibility of long-term drug testing with defined concentrations by adding the compounds directly to the medium. To validate the efficiency of a certain compound apoptosis rat

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