Culture of Bladder Cancer Organoids as Precision Medicine Tools.

Current in vitro therapeutic testing platforms lack relevance to tumor pathophysiology, typically employing cancer cell lines established as two-dimensional (2D) cultures on tissue culture plastic. There is a critical need for more representative models of tumor complexity that can accurately predict therapeutic response and sensitivity.

The development of three-dimensional (3D) ex vivo culture of patient-derived organoids (PDOs), derived from fresh tumor tissues, aims to address these shortcomings. Organoid cultures can be used as tumor surrogates in parallel to routine clinical management to inform therapeutic decisions by identifying potential effective interventions and indicating therapies that may be futile. Here, this procedure aims to describe strategies and a detailed step-by-step protocol to establish bladder cancer PDOs from fresh, viable clinical tissue. Our well-established, optimized protocols are practical to set up 3D cultures for experiments using limited and diverse starting material directly from patients or patient-derived xenograft (PDX) tumor material. This procedure can also be employed by most laboratories equipped with standard tissue culture equipment. The organoids generated using this protocol can be used as ex vivo surrogates to understand both the molecular mechanisms underpinning urological cancer pathology and to evaluate treatments to inform clinical management.

Journal of visualized experiments : JoVE. 2021 Dec 28*** epublish ***

Patrick B Thomas, Mahasha P J Perera, Saeid Alinezhad, Andre Joshi, Paria Saadat, Clarissa Nicholls, Caitlin P Devonport, Alivia R Calabrese, Abby R Templeton, Jack R Wood, Nathan J Mackenzie, Penny L Jeffery, Ian Vela, Elizabeth D Williams

School of Biomedical Sciences, Faculty of Health, Queensland University of Technology (QUT) at Translational Research Institute; Queensland Bladder Cancer Initiative (QBCI); Centre for Personalised Analysis of Cancers (CPAC); Australian Prostate Cancer Research Centre - Queensland., School of Biomedical Sciences, Faculty of Health, Queensland University of Technology (QUT) at Translational Research Institute; Queensland Bladder Cancer Initiative (QBCI); Centre for Personalised Analysis of Cancers (CPAC); Australian Prostate Cancer Research Centre - Queensland; Department of Urology, Princess Alexandra Hospital., School of Biomedical Sciences, Faculty of Health, Queensland University of Technology (QUT) at Translational Research Institute; Australian Prostate Cancer Research Centre - Queensland., School of Biomedical Sciences, Faculty of Health, Queensland University of Technology (QUT) at Translational Research Institute; Australian Prostate Cancer Research Centre - Queensland; Department of Urology, Princess Alexandra Hospital., School of Biomedical Sciences, Faculty of Health, Queensland University of Technology (QUT) at Translational Research Institute; Queensland Bladder Cancer Initiative (QBCI)., School of Biomedical Sciences, Faculty of Health, Queensland University of Technology (QUT) at Translational Research Institute; Queensland Bladder Cancer Initiative (QBCI); Australian Prostate Cancer Research Centre - Queensland., School of Biomedical Sciences, Faculty of Health, Queensland University of Technology (QUT) at Translational Research Institute; Queensland Bladder Cancer Initiative (QBCI); Centre for Personalised Analysis of Cancers (CPAC)., School of Biomedical Sciences, Faculty of Health, Queensland University of Technology (QUT) at Translational Research Institute; Queensland Bladder Cancer Initiative (QBCI); Centre for Personalised Analysis of Cancers (CPAC); Australian Prostate Cancer Research Centre - Queensland; .

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