Project description:Single-cell RNA-seq of engineered healthy human kidney organoids. These data are part of a larger investigation (data not provided here) showing that SARS-CoV-2 can directly infect engineered human blood vessel organoids and human kidney organoids which can be inhibited by human recombinant soluble ACE2 (hrsACE2), demonstrating that hrsACE2 can significantly block early stages of SARS-CoV-2 infections.
Project description:We used scRNA-Seq (and other biological experiments) to study the impact of a high oscillatory glucose regime in the differentiation and metabolic status of human kidney organoids further emulating early hallmarks of the diabetic kidney. To study the interplay between those changes and SARS-CoV-2 infection we assessed the effect of early stages of infection (1 day post infection) in normoglicemic (5 mM), hyperglicemic (5-25 mM oscillatory regime) and differentiating (11 mM) conditions. We retrieved metabolic changes and an upregulation of inflammatory related processes upon infection, whilst there was not an impact in the viability of the different cell types in the kidney organoids. Also, we studied the differences between ACE2 CRISPR/Cas9 knockouts and ACE2 WT organoids, using single cell RNA sequencing to resolve cell-type specific and cell proportion changes.
Project description: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.
Project description:These files represent single cell RNA-Seq data generated on a 10x Chromium genomics platform from four biological replicates of iPSC-derived human kidney organoids, in two batches, differentiated according to our published protocol (Takasato et al., Nature Protocols 2016). The aggregated human organoid data contains populations representing endothelial cells, podocytes, stroma, nephron, and off-target populations with similarity to neurons.
Project description:To reveal the key pathways involved in kidney cystogenesis. We performed single nuclei gene expression and ATAC profiling using kidney organoids from ARPKD patients-iPSCs derived organoids.