Urothelial organoids originate from mouse cells display differentiation capacity

Here we describe the establishment of mouse urothelial organoids from healthy urothelial tissue digests. NMU-o can grow within 7 days in medium enriched in growth factors, without FACS-sorting epithelial populations and can be passaged in culture for more than one year with stable morphology. In the absence of growth factors, NMU-o can differentiate within 14days exhibiting drastic morphological changes (increased diameter and lumen size, and decreased thickness layers) and expressing suprabasal urothelial markers (uroplakins and tigh junctions) both at the mRNA and protein levels. Transcriptomic tools (RNA-seq and single-cell RNA-seq) were preformed further supporting this differences between proliferative and differentiated organoids, allowing the discovery of novel genes involved in these processes.