PURPOSE 3D organotypic cell cultures offer the possibility to study cell growth in a more in vivo like situation. To our knowledge no 3D culture of primary urothelial cells has been established yet. BK Polyomavirus (BKPyV), replicating in urothelial cells, may cause haemorrhagic cystitis in immunocompromised patients. PRIMARY ENDPOINTS OF THIS STUDY Establishment of a 3D organotypic cell culture of primary urothelial cells and fibroblasts; use of this model as infection model for archetype BKPyV; description of first parts of viral life cycle with identification of therapeutic targets. METHODS This is an experimental study. Primary urothelial cells were purchased from CellnTec, Bern, Switzerland; fibroblasts were isolated from the ureter of patients with no urothelial malignancy in their medical history. As main methods we used quantitative real-time PCR and immunohistochemistry. Outcomes were analysed using SPSS 23.0. RESULTS We were able to develop a 3D organotypic culture for primary urothelium. An infection with archetype BKPyV was established in this model with virus replication rates up to 6.41 × 108 copies/ml on day 9 following Infection. Interestingly, proliferation rate of the urothelial cells is significantly (p = 0.049 at day 6 following infection) elevated while cells are losing differentiation under infection. Phosphorylated STAT3 is also significantly elevated (p < 0.0001) during infection. CONCLUSIONS The established of urothelial 3D cultures is a new method to study several urothelial diseases. The archetype BKPyV infection model is novel and the first method to study archetype viral life cycle. The STAT3 pathway might be an interesting target for the development of a causal therapy.

中文翻译：

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新型3D器官尿路上皮细胞培养模型，用于鉴定泌尿科感染的新治疗方法。

用途3D有机型细胞培养物提供了在更像体内的情况下研究细胞生长的可能性。据我们所知，尚未建立原代尿道上皮细胞的3D培养。在尿路上皮细胞中复制的BK多瘤病毒（BKPyV）可能会在免疫功能低下的患者中引起出血性膀胱炎。这项研究的主要终点建立了原代尿道上皮细胞和成纤维细胞的3D有机型细胞培养物。使用该模型作为原型BKPyV的感染模型；病毒生命周期第一部分的描述以及治疗靶点的描述。方法这是一项实验研究。尿路上皮原代细胞购自瑞士伯尔尼CellnTec；从他们的病史中没有尿路上皮恶性肿瘤的患者的输尿管中分离出成纤维细胞。作为主要方法，我们使用了定量实时PCR和免疫组化技术。使用SPSS 23.0分析结果。结果我们能够开发用于原发性尿路上皮的3D有机型培养。在此模型中建立了原型BKPyV感染，感染后第9天病毒复制率高达6.41×108拷贝/ ml。有趣的是，尿路上皮细胞的增殖速率显着提高（感染后第6天p = 0.049），而细胞在感染下却失去了分化。感染期间磷酸化的STAT3也显着升高（p <0.0001）。结论建立尿道上皮3D文化是研究几种尿道上皮疾病的新方法。原型BKPyV感染模型是新颖的，是研究原型病毒生命周期的第一种方法。