Advanced rpmi 1 glutamax pen strep

hPSC stocks were maintained in mTeSR1 medium with daily medium changes and weekly passaging using Accutase or ReLeSR (STEMCELL Technologies, Vancouver). For differentiation into organoids, iPSCs (WTC-11 cell line, Coriell # GM25256) bearing knockout mutations in PKD2 were plated at 2,000 cells/well in 24-well plates, or 200 cells/well in 384-well plates, pre-coated with 300 μl of DMEM-F12 containing 0.2 mg/ml Matrigel and sandwiched the following day with 0.2 mg/ml Matrigel in 500 μl of mTeSR1 (STEMCELL Technologies, Vancouver) to produce scattered, isolated spheroid colonies. Coating and plating of 384-well plates was performed using a Matrix Wellmate liquid handling robot. 48 h after sandwiching, hPSC-derived spheroids were treated with 12 μM CHIR99021 (Tocris Bioscience) for 36 h in 1,000 μl of Advanced RPMI + 1× Glutamax + Pen-strep (all from Thermo Fisher Scientific), then changed to RB (Advanced RPMI + 1× Glutamax + 1× B27 Supplement, all from Thermo Fisher Scientific). For full details see Supplementary Methods.