Kasman Laura, Voelkel-Johnson Christina
Department of Microbiology & Immunology, Medical University of South Carolina.
J Vis Exp. 2013 Dec 1(82):50181. doi: 10.3791/50181.
Bladder cancer is the second most common cancer of the urogenital tract and novel therapeutic approaches that can reduce recurrence and progression are needed. The tumor microenvironment can significantly influence tumor development and therapy response. It is therefore often desirable to grow tumor cells in the organ from which they originated. This protocol describes an orthotopic model of bladder cancer, in which MB49 murine bladder carcinoma cells are instilled into the bladder via catheterization. Successful tumor cell implantation in this model requires disruption of the protective glycosaminoglycan layer, which can be accomplished by physical or chemical means. In our protocol the bladder is treated with trypsin prior to cell instillation. Catheterization of the bladder can also be used to deliver therapeutics once the tumors are established. This protocol describes the delivery of an adenoviral construct that expresses a luciferase reporter gene. While our protocol has been optimized for short-term studies and focuses on gene delivery, the methodology of mouse bladder catheterization has broad applications.
膀胱癌是泌尿生殖道第二常见的癌症，因此需要能够减少复发和进展的新型治疗方法。肿瘤微环境可显著影响肿瘤发展和治疗反应。因此，通常希望在肿瘤细胞起源的器官中培养它们。本方案描述了一种膀胱癌原位模型，其中通过导管插入术将MB49小鼠膀胱癌细胞注入膀胱。在该模型中成功植入肿瘤细胞需要破坏保护性糖胺聚糖层，这可以通过物理或化学方法实现。在我们的方案中，在细胞注入前用胰蛋白酶处理膀胱。一旦肿瘤形成，膀胱导管插入术也可用于递送治疗药物。本方案描述了表达荧光素酶报告基因的腺病毒构建体的递送。虽然我们的方案已针对短期研究进行了优化，并侧重于基因递送，但小鼠膀胱导管插入术的方法具有广泛的应用。
