Figure 1.
Human RPE Cells Cultured in Miniaturized Format Express Mature RPE Markers Are Polarized and Functional
(A) Immunofluorescence images of hESC-RPE that are plated in 384-well plates at a density of 45,000 cells/well for 13 days. Cells express mature RPE markers including MITF and MERTK. They also show polarized expression pattern of ZO1 and MERTK as shown in the orthogonal views.
(B) POS were subjected to different sonication times with 10% intensity to obtain homogeneous size distribution before addition to the cells. Before sonication, big and small POS clumps were observed in Alexa Fluor 555 (AF555)-labeled POS. After sonication for 5 s, POS clumps appeared more uniform.
(C) Quantification of fluorescence images of AF555-labeled POS after sonication. Data are represented as the mean of the size of POS particles ± SD. Sonication for 5 s with 10% intensity was chosen to be used in the phagocytosis assay.
(D) Increasing concentrations of POS were added to hESC-RPE in 384-well plates. Data represent the mean of the total POS/cell count ±SD. N = 4 RPE differentiation batches (two wells per batch). Significance was calculated using one-way ANOVA comparing all samples with each other. Addition of 106 POS/well increases the number of total POS/cell significantly compared with the first concentration of 2 × 105.