Organ Cultures of Adult Porcine Retina: A Model for Studing Neuroprotection in the Retina

Abstract

Abstract: : Purpose: To establish an in vitro model of preserved retinal tissue architecture by using adult porcine retina based on its similarity with the human retina. Methods: Adult porcine eyes were obtained from a local slaughterhouse and retinas were removed within 1–3 hours post mortem under aseptic conditions. Explants were prepared from porcine retinas attached or detached from the underlying pigment epithelium with the ganglion cell layer (GCL) facing down or up. Explants were maintained from 1 day to 1 week in culture dishes with serum–supplemented medium. After this time, retinas were fixed and processed in order to perform immunohistological studies both in sections and in toto. We analyzed the structure of the different retinal layers and the viability of different retinal cell types in the explants. Results: After 1 day of culture, the appearance of the porcine retina histological was histologically similar to that of the control retina and its architecture was well preserved. A progressive degeneration of the retinal structure was observed from day 2 of culture, mainly when the explants were performed from detached retinas. A distinct pattern of degeneration was observed when explants were performed with the GCL facing down or up. Thus, when the GCL was facing up, the outer and inner nuclear layers became more irregular and Müller cells showed a larger hypertrophy than in explants with ganglion cells facing down. An increase in GFAP expression was observed in Müller cells, whereas anti–neurofilament immunocytochemistry revealed degeneration of retinal ganglion cells and horizontal cells. Changes in retinal structure were less evident in explants performed with the GCL facing down in spite of being detached from the pigmented epithelium. Conclusions: Organ culture of the adult porcine retina can be a useful tool for analyses requiring a preserved architecture of the retina and intact cell–cell associations, at least for short periods of time.

Keywords: retinal culture • retinal degenerations: cell biology • immunohistochemistry