Identification of cell surface markers and establishment of monolayer differentiation protocol for retinal pigment epithelial cells.

In vitro differentiation of human pluripotent stem cells into functional retinal pigment epithelial (RPE) cells for cell based reparative therapy of age-related macular degeneration (AMD). In the present study, we identify CD140b, CD56, GD2 and CD184 as central cell surface markers to evaluate hPSC-RPE differentiation efficiency, as well as a potential tool for the enrichment of hPSC-RPE during and after differentiation. Using these markers together with single cell RNA sequencing to evaluate the differentiation process, we have established an efficient xeno-free and defined monolayer differentiation methodology where culture on supportive human recombinant laminin eliminates the need for manual selection, allowing large-scale production of pure hPSC-RPE.