97th DOG Annual Meeting 1999
ADULT RETINA AND PIGMENT EPITHELIUM IN PERFUSION CULTURE - A NEW ORGANOTYPICAL IN VITRO MODEL FOR PHYSIOLOGICAL AND PHARMACO-TOXICOLOGICAL STUDIES
K. Kobuch, S. Kloth, W. Herrmann, E. Eckert, J. Roider, V. P. Gabel
Up to now cell culture systems are mainly based on proliferating single cell types or embryonic tissue, which both differ considerably from adult tissue concerning the degree of differentiation and celltyp specific functions. For that reason an organotypic in vitro model of adult retina and RPE in perfusion culture was developed, compared with static culture and evaluated as an in vitro toxicity model for vitreous substitutes.
Methods: Freshly dissected retinae with adjacent RPE and choroid were put in a special double ring tissue holder with a diameter of 13mm (Minusheet®). The ring was placed in a double chamber perfusion container and the system was constantly perfused with medium from both sides, retina and choroid, with a speed of 1ml/h. 36 normal retinae and 18 retinae after application of different vitreous substitutes (Sil.oil, Decalene, O62) were examined. After 4, 7 and 10 days of permanent perfusion the tissue was processed for histological examination. Results were compared with static cultures with and without vitreous substitutes.
Results: The retinal architecture was well preserved after 10 days of perfusion. The RPE was still a fully differentiated intact monolayer on Bruch´s membrane with apical position of pigment and well preserved intercellular junctions. The structural integrity of the retina after perfusion culture was significantly better than after static culture. In this in vitro model of the retina various morphological changes were induced by the different vitreous substitutes. These results correlated with in vivo studies on rabbit eyes.
Conclusions: Adult retina and RPE can be well preserved in perfusion culture for longer periods. This organotypical in vitro model offers an alternative for short term in vivo studies. Also in vitro studies on human retinal tissue are possible with this system. Other perspectives for the application of this model are toxicity teste on retina and RPE and retinal and RPE-transplantation.
University Eye Hospital, Franz-Josef-Strauss-Allee 11, G-93053 Regensburg