Optimized Retinal Cell Dissociation for Downstream Analysis of Cone Photoreceptors.

The isolation of viable and homogenous retinal cells is essential for downstream molecular analysis, such as single-cell sequencing and transcriptomics. However, the dissociation of retinal cells can be challenging due to the intricate nature of retinal tissue and the fragility of its cells. Furthermore, cone photoreceptors are highly susceptible to degeneration. We developed an optimized protocol for retinal cell dissociation that improves the quality of cones for downstream molecular analysis. Our protocol employs a papain enzymatic digestion, mechanical dissociation, and filtration to obtain a high percentage of viable cells with minimal cell death and damage. This protocol is a reliable and efficient method for downstream molecular analysis of cones and other retinal cells which can be useful for disease modeling-in which target cells are already sensitive and in a degenerative state-and examining the effects of treatments such as gene replacement therapies.