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Soile Nymark, Symantas Ragauskas, Virpi Savolainen, Andrea Holme, Jari AK Hyttinen, Giedrius Kalesnykas; Morphological and Functional Characterization of Ex Vivo Retinal Explants from Adult Mice. Invest. Ophthalmol. Vis. Sci. 2014;55(13):2377.
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© ARVO (1962-2015); The Authors (2016-present)
In order to facilitate the development of new therapies, in vitro model systems are needed to investigate the effects, benefits and risks of the treatments to the retinal tissue. In this study, we characterized a retinal ex vivo culture system from adult mice using immunohistochemistry, stereology and electrophysiology.
Retinae of young C57Bl/6J mice (age under 8 weeks) were isolated and cultured under ex vivo conditions up to 17 days in serum free medium supplemented with B27 and N2. Electrophysiological characterization of the retinae was performed by microelectrodearray (MEA) technique. Spontaneous activity of ganglion cells as well as their responses to flashes and steps of light was recorded during the culturing period. Viability of the tissue was also followed by immunohistochemistry and stereology.
Our immunohistochemical analysis shows that cultured retinal explants preserve retinal morphology for at least 17 days, although significant thinning of the outer nuclear layer was observed. Retinal functionality is retained substantially even though it is gradually lost during the whole culturing period. Spontaneous electrical activity of retinal ganglion cells was recorded up to 14 days in culture. The responsiveness of ganglion cells to light stimulation was lost faster, during the first week in culture.
Based on our electrophysiological and immunohistochemical characterization, retinal explants from adult mice retain their morphology and functionality for a remarkably long time period in optimal culturing conditions. This culture system thus represents a valuable tool for screening neuroprotective compounds using morphological and functional correlates ex vivo.
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