Single-cell RNA-seq of human kidney organoids [5_250K cells]
Human pluripotent stem cells (hPSCs) derived organoids hold great promise for transplantation purposes however technological advances to overcome their massive production while ensuring differentiation fidelity are still missing. Here we develop a new procedure sustaining the derivation of hPSC-kidney organoids to identify a new scalable, uniform, and affordable approach empowering differentiation and cellular diversity. Using single-cell RNA sequencing, confocal image analysis and metabolic assays we show that hPSC-kidney organoids exhibit transcriptional variety and cellular composition which is boosted forcing cell-to-cell contact. To demonstrate the applicability of these findings we infused CRIPSR/Cas9 engineered WT1 GFP kidney organoids during normothermic machine perfusion (NMP) using porcine kidneys. NMP flow patterns showed stable rates among biological experiments. Samples were analyzed by in situ hybridization, confocal microscopy, and flow cytometry identifying cells of human origin after infusion. This work paves the way to the utilization of hPSC-kidney organoids for ex vivo cell-based therapies to improve graft function before transplantation. Overall design: Single-cell RNA-seq of human kidney organoids