Enhancing human kidney organoid differentiation from pluripotent stem cells with high-throughput automation

Organoids derived from human pluripotent stem cells are a potentially powerful tool for high-throughput screening (HTS), but the complexity of organoid cultures poses a significant challenge for miniaturization and automation. In this study, single-cell RNA sequencing identify within organoids previously-undetected parietal and interstitial compartments. We discovered that addition of vascular endothelial growth factor (VEGF) during the differentiation process resulted in an approximately ten-fold increase in endothelial cells, without compromising the formation of the organoids. Although VEGF clearly increased the number of endothelial cells by immunofluorescence, relatively few endothelial cells were captured by scRNA-seq and only a modest increase in endothelial cells was observed. This suggested that either a substantial number of endothelial cells were lost or destroyed before sequencing, or that a spectrum of maturation states was present in the cultures. We have used DropSeq to perform single cell sequencing on human kidney organoids.Organoids included in this analysis were generated by treatment with and without VEGF. We did a bulk RNA analysis on the organoids treated with and without VEGF