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. Author manuscript; available in PMC: 2021 Sep 1.
Published in final edited form as: Dev Biol. 2021 May 15;477:98–116. doi: 10.1016/j.ydbio.2021.04.009

Fig. 1. hESC-derived kidney organoids display expected renal cell types.

Fig. 1.

A) Schematic and timeline of human embryonic stem cell (hESC) kidney organoid protocol. H9 ESCs were cultured in TeSR-E6 with 6 μM CHIR, with a switch to 200 ng/ml Fgf9 at day −3. On day 0, the differentiated cells were aggregated and placed into an ultra-low attachment 96-well plate with 200 ng/ml FGF9 for 5 days before being switched to growth-factor free media. B-D) Immunofluorescence of day 0 differentiated hESCs. 150 μm scale bars. B) Accumulations of cells with Meis1/2/3+ stroma surrounded by Six2+ NPCs. Cells in the middle forming tubule-like rings as seen by DAPI (arrows). C) Formation of E-Cad+ epithelial structures. D) VEGFR2+Sox17− angioblasts. E) Brightfield images of hESC organoids 2–7 days after aggregation. 0.5 mm scale bar. F-G) Immunofluorescence of day 12 hESC organoids demonstrating F) E-Cad+ epithelium and LTL+ proximal tubules and G) Podxl+ podocytes. H) Paraffin section of Meis1/2/3+ stroma. 100 μm scale bars. DAPI marks nuclei. N≥3 from at least 3 independent experiments.