Culture of human retinal explants as an ex vivo model for retinal gene therapy

Abstract

AbstractOptogenetic gene therapy may employ recombinant adeno-associated virus (AAV) vectors for specific preclinical applications in the treatment of inherited retinal diseases. Human retinal explants as an ex vivo model assist determine cellular tropism and the level of subsequent gene expression followed by delivery of viral vectors. Extracts of retinal tissue can be obtained and cultured for a certain time; thus, it is possible to transduce and confirm the effectiveness of the virus using various methods, including immunohistochemical staining. Consequently, we described method for harvesting and culturing human retinal fragments from divergent parts of the retina that differ anatomically and molecularly. The fragments were fixed, sectioned and stained. A dissociation method has also been described to obtain a retinal cell suspension that can be used in other experiments such as flow cytometry or mRNA/protein level expression assays.