2026-04-17T19:17:02Zhttps://recercat.cat/oai/request

oai:recercat.cat:10230/427512025-12-19T22:58:56Zcom_2072_6col_2072_452952

00925njm 22002777a 4500 dc Santos, Catarina P. author Lapi, Eleonora author Martínez de Villarreal, Jaime author Álvaro-Espinosa, Laura author Fernández Barral, Asunción author Barbáchano, Antonio author Domínguez, Orlando author Laughney, Ashley M. author Megías, Diego author Muñoz, Alberto author Real, Francisco X. author 2019-11-06T10:51:31Z 2019-11-06T10:51:31Z 2019 Understanding urothelial stem cell biology and differentiation has been limited by the lack of methods for their unlimited propagation. Here, we establish mouse urothelial organoids that can be maintained uninterruptedly for >1 year. Organoid growth is dependent on EGF and Wnt activators. High CD49f/ITGA6 expression features a subpopulation of organoid-forming cells expressing basal markers. Upon differentiation, multilayered organoids undergo reduced proliferation, decreased cell layer number, urothelial program activation, and acquisition of barrier function. Pharmacological modulation of PPARγ and EGFR promotes differentiation. RNA sequencing highlighted genesets enriched in proliferative organoids (i.e. ribosome) and transcriptional networks involved in differentiation, including expression of Wnt ligands and Notch components. Single-cell RNA sequencing (scRNA-Seq) analysis of the organoids revealed five clusters with distinct gene expression profiles. Together, with the use of γ-secretase inhibitors and scRNA-Seq, confirms that Notch signaling is required for differentiation. Urothelial organoids provide a powerful tool to study cell regeneration and differentiation. This work was supported, in part, by grants from Spanish Ministry of Economy, Industry and Competitiveness (SAF2016-76377-R), CIBERONC (CB16/12/00453 and CB16/12/00273), and Fundación Científica de la Asociación Española Contra el Cáncer. CNIO is supported by Ministerio de Ciencia, Innovación y Universidades as a Centro de Excelencia Severo Ochoa SEV-2015–0510. Bladder Differentiation Stem cells Urothelial organoids originating from Cd49fhigh mouse stem cells display Notch-dependent differentiation capacity