Fig. 1: hPSC-RPE cell surface marker screening and validation.

a, b Schematics of the antibody library screen and dot-plot graphs displaying the most relevant markers identified with the antibody library and their relative degree of expression between the hESC and optic vesicle (OV) cell populations (a) and between the hESC and day 60 hPSC-RPE populations (b). Each dot represents a different cell surface protein, and their position along the x and y axes is determined by the percent positive value in hESC and optic vesicle-cell/hPSC-RPE samples. Based on their position in the chart, a subset of cell surface proteins have been categorized as hPSC specific (bottom-right region) or optic vesicle specific (top-left region). c Flow cytometry histograms representing percentage of positive cells for CD140b, GD2, and CD184 in the pigmented and non-pigmented fractions of the embryoid bodies after 30 days of differentiation. Representative bright field pictures depicting the pigmented and non-pigmented fractions of the embryoid bodies that were analyzed by flow cytometry. Negative gates were set based on fluorescence minus one (FMO) control samples. Results are based on pooled samples from three independent differentiations. d Immunofluorescence stainings displaying the expression pattern of CD140b, CD184, and GD2 cell surface markers in day 60 hPSC-RPE cells. e Upper: Bright field and immunofluorescent pictures displaying the expression pattern of CD140b and human-specific BEST-1 (does not label rabbit BEST-1) in albino rabbit subretinally injected with hPSC-RPE cells. Pigmentation is of human origin as albino rabbits lack pigmentation. Lower: Bright field immunohistochemistry pictures showing the expression of CD140b in a human subretinal tissue section. f Bright field and immunofluorescent images showing pigmentation, as well as BEST-1 and CD140b co-expression patterns in the CD140b⁺ and CD140b⁻ populations sorted at day 30 of differentiation. g Bar graphs representing the quantification of cells that are pigmented, BEST-1⁺, CD140b⁺, BEST-1 and CD140b⁺ in the CD140b⁺ and CD140b⁻ sorted populations. Bars represent means ± SEM from four different images. Scale bars: c = 200 μm; d = 20 μm; e, f = 50 μm. Source data are provided as a Source Data file.