kidney organoids derived from human induced pluripotent stem cells (iPSC)

iPSCs were maintained on Geltrex coated flat culture dish in E8 media according to manufacturer’s guidelines. Colonies were manually harvested at 60-80% confluence. Cells were then collected and dissociated into single cells using EDTA. Cells were put on to ultra low attachment 24 well or 96 well plate to allow them to form aggregated in suspension with ROCK inhibitor. Cell aggregates 191were cultured in E8 medium with daily medium change for 6-7 days. Control iPSC-A (iPSC-aggregates) were plated on a Geltrex in 96 well plate or 8 well culture chamber. And then aggregates were treated E8 medium with daily medium change for 12-14 days. Starting from human IPSC in vitro kidney differentiation was performed in order to study transcriptome regulated program during this stage of development: initial human IPSC cell (PB33) was processed in duplicate et resulting kidney organoid was processed in duplicate