Mouse and human urothelial cancer organoids: A tool for bladder cancer research

Significance Bladder cancer poses a big clinical challenge, particularly in advanced cases. Here we present a culture system for bladder cancer based on organoid culture technology. We have created a living biobank containing organoids grown from over 50 patient samples, which reflects many aspects of bladder cancer pathogenesis. Organoids of human bladder cancer cells can be maintained for prolonged periods of time, and closely resemble the tumor histology. Bladder organoids can be used for drug screening, and therefore provide a platform for development of new drugs for the treatment of bladder cancer and personalized medicine. Bladder cancer is a common malignancy that has a relatively poor outcome. Lack of culture models for the bladder epithelium (urothelium) hampers the development of new therapeutics. Here we present a long-term culture system of the normal mouse urothelium and an efficient culture system of human bladder cancer cells. These so-called bladder (cancer) organoids consist of 3D structures of epithelial cells that recapitulate many aspects of the urothelium. Mouse bladder organoids can be cultured efficiently and genetically manipulated with ease, which was exemplified by creating genetic knockouts in the tumor suppressors Trp53 and Stag2. Human bladder cancer organoids can be derived efficiently from both resected tumors and biopsies and cultured and passaged for prolonged periods. We used this feature of human bladder organoids to create a living biobank consisting of bladder cancer organoids derived from 53 patients. Resulting organoids were characterized histologically and functionally. Organoid lines contained both basal and luminal bladder cancer subtypes based on immunohistochemistry and gene expression analysis. Common bladder cancer mutations like TP53 and FGFR3 were found in organoids in the biobank. Finally, we performed limited drug testing on organoids in the bladder cancer biobank.