Decellularized retinal matrix: Natural platforms for human retinal progenitor cell culture

Abstract

Tissue decellularization strategies have enabled engineering of scaffolds that preserve native extracellular matrix (ECM) structure and composition. In this study, we developed decellularized retina (decell-retina) thin films. We hypothesized that these films, mimicking the retina niche, would promote human retinal progenitor cell (hRPC) attachment, proliferation and differentiation. Retinas isolated from bovine eyes were decellularized using 1% w/v sodium dodecyl sulfate (SDS) and pepsin digested. The resulting decell-retina was biochemically assayed for composition and cast dried to develop thin films. Attachment, viability, morphology, proliferation and gene expression of hRPC cultured on the films were studied in vitro. Biochemical analyses of decell-retina compared to native retina indicated the bulk of DNA (94%) was removed, while the majority of sulfated GAGs (55%), collagen (83%), hyaluronic acid (87%), and key growth factors were retained. The decell-retina films supported hRPC attachment and growth, with cell number increasing 1.5-fold over a week. RT-PCR analysis revealed hRPC expression of rhodopsin, rod outer membrane, neural retina-specific leucine zipper neural and cone-rod homeobox gene on decell-retina films, indicating photoreceptor development. In conclusion, novel decell-retina films show promise as potential substrates for culture and/or transplantation of retinal progenitor cells to treat retinal degenerative disorders. Statement of significanceIn this study, we report the development of a novel biomaterial, based on decellularized retina (decell-retina) that mimics the retina niche and promotes human retinal progenitor cell (hRPC) attachment, proliferation and differentiation. We estimated, for the first time, the amounts of collagen I, GAGs and HA present in native retina, as well as the decell-retina. We demonstrated that retinas can be decellularized using ionic detergents and can be processed into mechanically stable thin films, which can act as substrates for culturing hRPCs. Rhodopsin, ROM1, NRL and CRX gene expression on the decell-retina films indicated photoreceptor development from RPCs. These results support the potential of decell-retina as a cell delivery platform to treat and manage retinal degenerative disease like AMD.