Fig. 2: Generating mature and highly organized CD organoids from UB organoids.

a Schematic of the screening strategy for identifying differentiation culture condition to generate CD organoids from UB organoids. b qRT-PCR analyses of UB (red) and CD (green) organoids for UB progenitor markers Wnt11 and Ret; PC markers Aqp2 and Aqp3; IC markers Foxi1, Atp6v1b1, Slc4a1, and Slc26a4; and Tfcp2l1 that is expressed in both PC and IC. Adult mouse kidney (blue) was used as control. The significance was determined by two-tailed unpaired Student’s t tests. n = 3. c Bright field images showing the morphologic differences between UB and CD organoids. d–g Whole-mount immunostaining analyses of CD organoids for PC marker AQP2, and IC markers FOXI1 and ATP6V1B1, showing the distribution of these two cell types within the organoid. f Shows the higher-power images for the boxed region in (e). h Comparison of PC and IC ratios in postnatal Day 0 (P0) mouse CD, adult mouse CD, and CD organoids. Whole-mount immunostaining images (CD organoids), or section staining images (P0 and adult kidneys) stained for AQP2 (PC) and FOXI1 (IC) were quantified for ratios of PC and IC in the CD organoid or the kidney’s collecting duct. Each column represents counts from nine different fields of view (n = 9, three fields were randomly selected from each of the three biologically independent samples). i Principal component analysis (PCA) of RNA-seq data. Different colors and oval circles represent different primary kidney cell populations (NPC, IPC, UB tip, UB trunk, and adult CD cells), or UB organoids cultured for 5 (D5), 10 (D10), and 20 days (D20), or CD organoids. Scale bars: c, d 200 µm, e 100 µm, f 25 µm, g 40 µm. All data are presented as mean ± s.d. Source data are provided as a Source Data file.