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      Roller Culture of Free-Floating Retinal Slices: A New System of Organotypic Cultures of Adult Rat Retina

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          No experimental system exists to date for the in vitro study of retinal ganglion cell populations in a three-dimensional organotypic tissue environment. Here, we describe such a novel method for roller cultivation of adult retinas. Retinas of adult (1–3 months old) rats were cut into rectangular slices of approximately 1 mm<sup>2</sup>. Free-floating slices were cultured on a horizontal rotating roller drum (50–60 rpm) in a dry incubator at 36.5°C. During the first days of cultivation, primary flat retinal slices changed their configuration and transformed into ball-shaped tissue spheres (retinal bodies). Histological and immunocytochemical studies showed that the outer wall of the retinal bodies was formed by cell and fibre layers typical of mature retina with photoreceptors located on the outside. Initially, retinal bodies contained an inner cavity which later was completely obliterated and filled with glial cells, sprouting nerve fibres, and vascular structures. This culture system was further developed into a robust model of glutamate-induced neurotoxicity. Using a novel culture method of adult rat retina, preservation of the three-dimensional organotypic retinal cytoarchitecture was achieved, including survival of neurons in the ganglion cell layer and sprouting of nerve fibres of the axotomized retinal ganglion cells. This novel culture model promises to facilitate studies of retinal physiology and pathology.

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          Most cited references 31

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          Ganglion cell death in glaucoma: what do we really know?

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            A reliable method for organ culture of neonatal mouse retina with long-term survival.

            Organ culture systems of the central nervous system have proven to be useful tools for the study of development, differentiation, and degeneration. Some studies have been limited by the inability to maintain the cultures over an extended period. Here we describe an organ culture technique for the mouse retina. This method uses commercially available supplies and reproducible procedures to maintain healthy retinas with normal architecture for 4 weeks in vitro. The system is amenable to quantitative analysis. It can be used with both normal and retinal degeneration (rd) retinas to study of the role of various factors in photoreceptor degeneration in retinal cell fate determination and development.
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              Mouse retina explants after long-term culture in serum free medium


                Author and article information

                Ophthalmic Res
                Ophthalmic Research
                S. Karger AG
                October 2006
                03 October 2006
                : 38
                : 5
                : 263-269
                aLaboratory of Experimental Neurocytology, Brain Research Institute, Moscow, Russia; bDepartment of Neurology, Experimental Neurology, Campus Charité Mitte, cDepartment of Psychiatry, Campus Benjamin Franklin, and dDepartment of Psychiatry, Campus Charité Mitte, Berlin, Germany
                95768 Ophthalmic Res 2006;38:263–269
                © 2006 S. Karger AG, Basel

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                Page count
                Figures: 3, References: 39, Pages: 7
                Original Paper


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