Kidney Organoid Differentiation from hiPSCs

Healthy control (WTC-11), GITpatient, and GITpatient^corr hiPSCs were differentiated into kidney organoids following the previously published protocol [10 (link),11 (link),12 (link),14 (link)] (Figure 3B). Briefly, hiPSCs were plated in mTeSR1 medium (05850; STEMCELL Technologies Vancouver, Canada) supplemented with 10 μM Y-27632 (1293823; Biogems, Westlake Village, CA, USA) onto 24-well plates pre-coated with 1.25% Corning Matrigel^® hESC Qualified Matrix. After 24 h, the medium was exchanged with 2.5% of Matrigel^® in mTeSR1. On the fourth day, the medium was replaced with Advanced RPMI (1263302; Thermo Fisher Scientific, Grand Island, NY, USA) supplemented with 12 µM CHIR-99021 (STEMCELL Technologies). Approximately 36 h later, the medium was changed to Advanced RPMI with B27 supplement (17504044, Thermo Fisher Scientific). Organoids were cultured in this medium until collection on day 21.