Optimizing Kidney Organoid Response

To determine optimal concentration of each compound, PKHD1-mutant organoids were treated with the following concentration from day 16 to day 35 of differentiation: T-5224 (Thermo Fisher Scientific, 50-115-1835) at 5, 10, and 20 μM; Z-DEVD-FMK at 1, 10, and 50 μM (R&D Systems, FMK004); quercetagetin at 10, 40, and 200 μM (Millipore Sigma, 551590); NSC23766 at 25, 50, and 100 μM (Abcam, ab142161); rhCXCL16 at 5, 10, and 25 ng/ml (R&D Systems, 976-CX-025); 2-MeOE₂ at 1, 5, and 10 μM (Selleckchem, S1233); R-naproxen at 10, 20, 200, and 400 μM (Millipore Sigma, 82170); and R-ketorolac at 0.5, 1, 10, and 20 μM (Millipore Sigma, 1654) (45 (link), 46 (link), 73 (link)–77 (link)). Full medium exchanges were conducted every 2 to 3 days. Treatment of the kidney organoids-on-a-chip was started at day 16 of their differentiation until day 35. The compounds were added to the medium reservoir at every medium change in the following concentrations: T-5224 at 10 μM, Z-DEVD-FMK at 1 μM, quercetagetin at 10 μM, NSC23766 at 10 μM, rhCXCL16 at 1 ng/ml, 2-MeOE₂ at 5 μM, R-naproxen at 20 μM, S-naproxen at 20 μM, and R-ketorolac at 1 and 10 μM.