Fig. 1: HRO system reproducing several characteristics of the human retina. | Nature Communications

Fig. 1: HRO system reproducing several characteristics of the human retina.

From: HBEGF-TNF induce a complex outer retinal pathology with photoreceptor cell extrusion in human organoids

Fig. 1

a Schematic. HRO generation, retinal structure, and cell types quantified on (b, c) immunostained serial HRO sections: cones (ARR3); rods (NRL); MG (SOX9, RLBP1). c Graphs: circles represent individual HROs (n) derived from N = 4 independent experiments, n ≥ 5/N; mean ± SD over n. Part of the dataset presented in Fig. 3c, Supplementary Data 2. d Voronoi neighbor analysis: number of cone neighbors analyzed from en-face images of HRO wholemounts immunostained for cone (ARR3), photoreceptor (RCVRN), and Müller glia (SLC1A3 + RLBP1) markers (Supplementary Fig. 3a–e, k; N = 1; n = 8; mean ± SD over n; 3 ROIs averaged/n). e Scanning electron microscopy of HRO indicated the formation of photoreceptor inner (PIS) and outer (POS) segments (N = 1, n = 6). f Rod (RHO) and cone (OPN1LW/MW, OPN1SW) opsin immunostaining on HRO sections (N = 1; n = 9). g HRO single-cell RNA-seq analysis: dotplot shows cell clusters after Louvain cluster detection in a UMAP embedding. Piechart: relative abundance of major cell types in an individual HRO. Dotplot/piechart: pseudocolored for cell types as indicated. Photoreceptors (light pink): likely immature cones, but could not be clearly assigned. h Comparison of single-cell RNA-seq data: distribution of Pearson correlation of HRO cells and human retinal fovea and periphery reference datasets58. Violin graph: Pearson correlation of cones, rods, and Müller glia in HRO (N = 1) against the human reference vectors (correlation values of three individual human donors (N = 3) combined in one violin graph; Supplementary Fig. 3g: individual donor data). Boxplots: minimum, 1st quantile, median, 3rd quantile, maximum values, dots are outlier data points (lower range: the 25th percentile-1.5 × IQR; upper range: 75th percentile + 1.5 × IQR; IQR: interquartile range). i The human anatomical macula: eye fundus image (at age 28) and schematic of the retina. Graph: macular cone–rod densities in humans in vivo (references: Supplementary Fig. 3i, j). Arrows: cone–rod ratio replicated in HROs; prevalent site of AMD/MDDs onset9,110, (Supplementary Fig. 3i, j). Scale bars: b 1 mm, c 50 µm, e 10 µm (inset 1 μm), f 10 µm. Source data are provided as a Source Data file.

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