Kim Yubin, Lee Juhee, Kim SungEun, Shin Kunyoo
Department of Life Sciences, Pohang University of Science and Technology.
Department of Life Sciences, Pohang University of Science and Technology;
J Vis Exp. 2020 Jan 31(155). doi: 10.3791/60469.
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.
长期以来，先进肿瘤模型的开发一直受到鼓励，因为目前的癌症模型已显示出局限性，如缺乏三维（3D）肿瘤结构且与人类癌症的相关性较低。研究人员最近开发了一种称为肿瘤类器官的3D体外癌症模型，它可以在培养皿中模拟天然肿瘤的特征。在此，详细描述了从致癌物诱导的小鼠膀胱肿瘤建立膀胱肿瘤类器官的实验程序，包括所得3D肿瘤类器官的培养、传代和体外维持。此外，还描述了使用慢病毒介导的转导对已建立的膀胱肿瘤类器官系进行基因工程操作的方案，包括将新的遗传元件有效引入肿瘤类器官的优化条件。最后，阐述了将膀胱肿瘤类器官原位移植到小鼠膀胱壁进行进一步分析的程序。本文所述方法有助于建立用于膀胱癌的体外模型，以开发更好的治疗方案。
