Abstract
The development of advanced tumor models has long been encouraged because current cancer models have shown limitations such as lack of three-dimensional (3D) tumor architecture and low relevance to human cancer. Researchers have recently developed a 3D in vitro cancer model referred to as tumor organoids that can mimic the characteristics of a native tumor in a culture dish. Here, experimental procedures are described in detail for the establishment of bladder tumor organoids from a carcinogen-induced murine bladder tumor, including culture, passage, and maintenance of the resulting 3D tumor organoids in vitro. In addition, protocols to manipulate the established bladder tumor organoid lines for genetic engineering using lentivirus-mediated transduction are described, including optimized conditions for the efficient introduction of new genetic elements into tumor organoids. Finally, the procedure for orthotopic transplantation of bladder tumor organoids into the wall of the murine bladder for further analysis is laid out. The methods described in this article can facilitate the establishment of an in vitro model for bladder cancer for the development of better therapeutic options.
Highlights
Bladder cancer is the most prevalent urinary tract cancer, with approximately 165,000 patients dying annually[1]
Favorable results have been observed in numerous drug candidates in cancer cell lines, poor results are reported in clinical trials[4]
NOTE: Passage of bladder tumor organoids when they reach 100–150 μm in diameter is recommended
Summary
Bladder cancer is the most prevalent urinary tract cancer, with approximately 165,000 patients dying annually[1]. Animal cancer models or patient-derived tumor xenografts can be used to address the limitations observed in bladder cancer cell lines. Over the last several years, tumor organoids have been established from a large number of diverse tumor tissues[7], including colon[8,9], bladder[10], pancreas[11,12], prostate[13], liver[14], and breast[15] tumor tissues. Such tumor organoids mimic their original tumors phenotypically and genetically. All procedures were approved and conducted under the guidelines of the Institutional Animal Care and Use Committee at POSTECH (IACUC number: POSTECH-2019-0055)
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