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  • Cross-platform single cell ...
    Magella, Bliss; Adam, Mike; Potter, Andrew S.; Venkatasubramanian, Meenakshi; Chetal, Kashish; Hay, Stuart B.; Salomonis, Nathan; Potter, S. Steven

    Developmental biology, 02/2018, Letnik: 434, Številka: 1
    Journal Article

    The developing kidney provides a useful model for study of the principles of organogenesis. In this report we use three independent platforms, Drop-Seq, Chromium 10x Genomics and Fluidigm C1, to carry out single cell RNA-Seq (scRNA-Seq) analysis of the E14.5 mouse kidney. Using the software AltAnalyze, in conjunction with the unsupervised approach ICGS, we were unable to identify and confirm the presence of 16 distinct cell populations during this stage of active nephrogenesis. Using a novel integrative supervised computational strategy, we were able to successfully harmonize and compare the cell profiles across all three technological platforms. Analysis of possible cross compartment receptor/ligand interactions identified the nephrogenic zone stroma as a source of GDNF. This was unexpected because the cap mesenchyme nephron progenitors had been thought to be the sole source of GDNF, which is a key driver of branching morphogenesis of the collecting duct system. The expression of Gdnf by stromal cells was validated in several ways, including Gdnf in situ hybridization combined with immunohistochemistry for SIX2, and marker of nephron progenitors, and MEIS1, a marker of stromal cells. Finally, the single cell gene expression profiles generated in this study confirmed and extended previous work showing the presence of multilineage priming during kidney development. Nephron progenitors showed stochastic expression of genes associated with multiple potential differentiation lineages. •Single cell RNA-seq of over 8000 E14.5 developing kidney cells.•Stromal cells, as well as cap mesenchyme, express GDNF.•Comparison of 10X Genomics, Drop-Seq and Fluidigm 800 cell IFC single cell RNA-Seq.•Multilineage priming at many stages of nephrogenesis.