Objective: To investigate the methods of isolation and culture of fetal human retinal photoreceptors.
Methods: Fetal human retinal photoreceptors were obtained by enzyme gradation digestion, cultured in media conditioned by retinal pigment epithelial cells (RPE-CM), identified by morphologic observation and immunohistochemically stained with rod specific opsin 4D2 (Rho 4D2) antibody. The digested retinal tissues were tested by hematoxylin-eosin (HE) staining.
Results: Enzyme gradation digestion could obtain relatively pure retinal photoreceptors. These cells were stained positively for Rho 4D2 antibody, and survived in RPE-CM for a long time.
Conclusion: Fetal human retinal photoreceptors obtained by enzyme gradation digestion can survive in RPE-CM for a long time and express rod specific opsin. This culture system will provide a source for human retinal photoreceptor transplantation and further studies of photoreceptors in vitro.