Gene Delivery using Adenoviral Vector into the Organotypic Culture System of the Chick Retinae

By: Mohamed Algriw , Abdulhakem Elnfati

Faculty of Science - University of Tripoli, Libya

Issue: Vol 20 |First Issue | 2014

article language: English

Abstract:

Damage to retina during development correlates with many ophthalmological pathologies, such as retinal dysplasia and degeneration. Virus-mediated gene transfer into identified retinal cells may offer a great potential for therapeutic interventions. This study describes a three-dimensional culture system of the embryonic chick retina. Samples from 17 and 19 days old retinae were isolated from eyeball and were cultured for up to 10 days. Selection of embryonic stage and specimen processing were optimized for culturing. The ability of a replication-deficient adenovirus vector (AVV) carrying an early cytomegalovirus (CMV) gene promoter to efficiently transfer a green fluorescent protein (GFP) gene into the cultured embryonic chick retinae was analysed. The results indicated that retinal tissue slices did not show any change in their cytoarchitecture. The results also showed that the GFP expression was maximal about 24 hours after infection of retinal slices and were maintained for several days. They also showed that GFP expression was widespread in the retinal slices. Moreover, the results also showed that injecting of AVV did not show any detectable cytopathic effects - these were only observed for very high titres of infection. The approach developed here is a valuable tool that can be used to assess the effect of pharmacological manipulations on retinal cells and to transfer of effective therapeutic genes into the retina, which might also facilitate the search for understanding the mechanisms responsible to vision-related diseases and to identify potential therapeutic interventions.