Fig. 1
From: 3D organoid-derived human glomeruli for personalised podocyte disease modelling and drug screening
Intact glomeruli can be isolated from human iPSC-derived kidney organoids. a Top left: Pure populations of whole glomeruli can be isolated from kidney organoids at scale by sieving. Scale bar 200 µm. Right: These organoid-derived glomeruli (OrgGloms) when examined in detail (scale bar 100 µm) show similarity to adult human glomeruli (Bottom left, scale bar 100 µm). b OrgGloms are formed in the appropriate size range, comparable to both human glomeruli at 32 weeks gestation70, and adult human glomeruli62. One-way ANOVA p < 0.0001; error bars = SEM; significant difference assessed by Tukey’s multiple comparisons test; F-value = 176.9; DF = 2; biological replicates n = 30 adult glomeruli, n = 30 neonatal glomeruli, n = 155 OrgGlom. c Glomeruli developed within kidney organoids show comparable morphology to capillary loop stage human glomeruli, highlighted by in situ immunofluorescent staining of serial sections with the podocyte protein NEPHRIN, which shows infolding of the glomerular surface layer. Scale bar 10 µm, central cross-section (left), surface (right). d Quantitative PCR analysis of isolated glomerular fractions compared to whole organoid show a significant enrichment in NPHS1 gene expression in sieved glomeruli. Error bar = SEM; significant difference assessed by Student’s unpaired t-test p = 0.0002, t-value = 13, DF = 4, n = 3 biological replicates shown by symbols. e Transmission electron microscopy of kidney organoid glomeruli show podocyte cell bodies connected by primary and secondary processes (inset), surrounded by an outer layer of parietal epithelial cells (arrowhead). Scale bar 10 µm. f Immunostaining of whole OrgGloms shows appropriate apicobasal cell polarity. A single cross-sectional plane is shown on the left, and a 3D reconstruction using all Z-stack images acquired (444 sections) on the right. Expression levels of single channels shown in greyscale to preserve maximum contrast, merged images shown in colour. Full 3D reconstruction video of the Z-stack can be found as Supplementary Video 1
