Fig. 1: hiPSC-derived kidney organoids source pseudo proximal tubule cells.

a The simplified protocol of hiPSC differentiation into intermediate mesoderm and formation of 3D kidney organoids, followed by dissociation, sorting, and seeding the cells as sorted. Dissociated organoid, LTL + and LTL–, refer to dissociated organoid cells, positive fraction of MACS, and negative fraction of MACS, respectively. b Select brightfield and confocal fluorescent images (z-intensity projected) of kidney organoids on day 22. Immunochemistry is for EpCAM and proximal tubule markers, LTL and megalin. c Effect of LTL concentration (dilution factors of 1/50 or 1/100) on the specificity of MACS, N = 4 experiments and the error bars represent standard deviation of data, *p < 0.05. d Relative gene expression levels obtained from cells as sorted and cultured for 7 days in culture dishes and PToC: Organoid, dissociated organoid cells on day 22; LTL+/–, positive/negative fraction of MACS products. White area corresponds to samples obtained from as-dissociated (organoid) and as-sorted cells, whereas the grayed area indicates samples from cells cultured for 7 days. Error bars represent the standard deviation with N = 3 biologically independent experiments. N.S., not significant for p > 0.05; *p ≤ 0.05; ** p ≤ 0.01; ***p ≤ 0.001; ****p ≤ 0.0001. e Evolution of LTL+ cells cultured on the membrane into aggregates, Scale bar, 200 μm. The yellow frame shows the boundary between the cell aggregate and the monolayer. Scale bar, 200 μm. f Immunochemistry on day 7 for EpCAM, LTL, and megalin, markers of proximal tubules, in sorted cells extracted from kidney organoids and seeded on the PToC. For the LTL+ tissue fluorescent scans were conducted on select parts at the proximity of aggregates as framed in (e). Scale bar, 50 μm.