Figure 6. Modeling kidney development and injury in kidney organoids.
(a) Schematic for kidney development analysis and nephrotoxicity assay. (b) Representative images of immunohistochemistry in structures derived from hESCs treated with DAPT 10 µM from day 14 to 21. n=4. Notch inhibition suppressed proximal tubule formation. Scale bars: 50 µm. (c) Representative immunohistochemistry in structures treated with gentamicin (5 mg/ml) from day 21 to 23 or cisplatin (5 µM) from day 21 to 22. n=6. Scale bars: 50 µm. Gentamicin and cisplatin induced the upregulation of KIM-1, and cisplatin suppressed CDH1 expression. (d) A low magnification image of gentamicin-treated organoids. Scale bar: 100 µm. CDH1: Cadherin-1 (E-cadherin). PODXL: Podocalyxin-like (Podocalyxin). LTL: lotus tetragonolobus lectin. KIM-1: kidney injury molecule-1. (e) Real-time quantitative PCR of KIM-1 in kidney organoids treated with gentamicin at indicated doses. Data is expressed as mean +/− SEM (n=10). (f) Representative immunohistochemistry of organoids treated with cisplatin (5 or 50 µM) from day 23 to 24 (24 hours). n=4. Scale bars: 50 µm.