Roller Organotypic Cultures of Postnatal Rat Retina

Viktorov, I. V.; Lyzhin, A. A.; Aleksandrova, O. P.; Frayer, David W.; Dirnagl, Ulrich

doi:10.1023/b:bebm.0000035147.97443.da

Cited by 7 publications

(9 citation statements)

References 10 publications

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“…Retinal body formation and development is similar in adult as compared with postnatal rat retina roller cultures [23] . However, some essential differences in the structure of the retinal bodies exist which are determined by differences in the level of retinal development and maturity at the beginning of culture.…”

Section: Discussionmentioning

confidence: 87%

“…The retina of 7-to 8-day-old rats is still developing, in spite of its cell-and-layer architecture which is similar to that of the adult rat retina. Development of the isolated postnatal retina continues during roller culture, and this leads to some structural characteristics which do not develop in roller cultures of adult rat retinas, e.g., formation of multilayered retinal bodies, confl uence of several retinal bodies, and rosette formation in the ONL [23] . A number of studies described survival of RGCs and axonal outgrowth in fl at-mounted organotypic tissue cultures of the adult rodent and human retina [27][28][29][30][31] .…”

Section: Discussionmentioning

confidence: 99%

“…In the present study, a high-speed roller drum method [22] , adapted initially for the culture of free-fl oating retinal slices of postnatal rats [23] , is described and characterized specifi cally for the organotypic cultivation of adult rat retinas, with survival and possibly regeneration of RGCs.…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2006

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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 mm². 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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Section: Discussionmentioning

confidence: 87%

Section: Discussionmentioning

confidence: 99%

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

et al. 2006

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“…After dehydration in ethanol (70%) and three portions of dehydrated (99.9%) isopropyl alcohol (Sigma) the retinas were placed into melted (58 o C) paraffin (Sigma) for 3 h or overnight [1]. After 1-h incubation in a new portion of paraffin the retinas were embedded in blocks.…”

Section: Methodsmentioning

confidence: 99%

“…Address for correspon dence: ivic@4unet.ru. I. V. ViktorovOrganotypic culturing of animal retina sections, apart from multilayer cell cultures, is used for the studies of the development and experimental diseases of mammalian retina [1,4,5,10]. The first organ cultures of the whole retina embedded in a plasma clot were obtained in 1930s [9].…”

mentioning

confidence: 99%

Roller organ cultures of the retina from postnatal rats

2006

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Whole retinas of 2-14-day-old rats were cultured in a roller device for 2-14 days. Floating retinas of 7-14-day-old rats formed hole spheroid structures (spheroids) with the wall completely retaining the linear structure and layer-by-layer cellular and fibrous architecture, including the outer nuclear, outer plexiform, inner nuclear, inner plexiform layers, layers of ganglion cells and nerve fibers. The retina obtained at earlier terms of development often formed folds, with pyknotic nuclei of dead neurons in their deep compartments. In organ cultures of the retina isolated from rats at early postnatal periods, rosettes were formed in sites of local injury to the outer nuclear layer and pigmented epithelium. Roller organ cultures can be used for in vitro studies of the development and experimental diseases of the retina.

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