Enhanced metanephric specification to functional proximal tubule enables toxicity screening and infectious disease modelling in kidney organoids

Improved specification of metanephric nephron progenitors in conditions that prolong differentiation while simultaneously preventing spontaneous nephron induction resulted in proximal tubule (PT)-enhanced kidney organoids. PT-enhanced organoids exhibited improved PT maturation, with elongated and aligned nephron segments as well as distinct loops of Henle. The striking proximo-distal orientation of nephrons was shown to result from localised WNT antagonism originating from the centre of the organoid. Overall design: Single cell RNA-seq generated on a 10x Chromium genomics platform from human iPSC-derived kidney differentiations and organoids. D13+14 sample represents 4 barcoded replicates of 4 pooled organoids, each replicate derived from seperate monolayer differentiations of CRL1502.2 iPSCs (D13 sample; also containing 4 barcoded replicates). D13+12 represents a single organoid derived from a D13 monolayer differentiation of CRL-2429/SIX2Cre/Cre:GAPDHdual iPSCs.