An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Francisco X. Real
Spanish National Cancer Research Centre-CNIO
Corresponding Author
ORCiD: https://orcid.org/0000-0001-9501-498X
License:
This work is licensed under a CC BY 4.0 License. Read Full License
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.
Keywords
Urothelium, organoids, stem cells, barrier function, differentiation, tight junctions, Cd49f/Itga6, PPARg, EGFR, Notch, Wnt, single-cell RNA Sequencing
Figure 1
The authors declare no competing financial interests
Loading...
Version History
CURRENT STATUS: Posted
Version 1
Posted 20 Nov, 2019
No community comments so far
Metrics
Comments: 0
HTML Views: ...
Associated Publications
Urothelial organoids originating from Cd49fhigh mouse stem cells display Notch-dependent differentiation capacity
by Catarina P. Santos, Eleonora Lapi, Jaime Martínez de Villarreal, +8
Nature Communications (27 September, 2019)
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Francisco X. Real
Spanish National Cancer Research Centre-CNIO
Corresponding Author
ORCiD: https://orcid.org/0000-0001-9501-498X
Version History
CURRENT STATUS: Posted
Version 1
Posted 20 Nov, 2019
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
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.
Figure 1
The authors declare no competing financial interests
Loading...
Associated Publications
Urothelial organoids originating from Cd49fhigh mouse stem cells display Notch-dependent differentiation capacity
by Catarina P. Santos, Eleonora Lapi, Jaime Martínez de Villarreal, +8
Nature Communications (27 September, 2019)