Chen Shuang, Giannakou Andreas, Golas Jonathon, Geles Kenneth G
Oncology R&D group, Pfizer Worldwide Research and Development.
Oncology R&D group, Pfizer Worldwide Research and Development;
J Vis Exp. 2020 Mar 17(157). doi: 10.3791/60644.
Tumor-stroma interactions play a critical role in the development of lung squamous carcinoma (LUSC). However, understanding how these dynamic interactions contribute to tissue architectural changes observed during tumorigenesis remains challenging due to the lack of appropriate models. In this protocol, we describe the generation of a 3D coculture model using a LUSC primary cell culture known as TUM622. TUM622 cells were established from a LUSC patient-derived xenograft (PDX) and have the unique property to form acinar-like structures when seeded in a basement membrane matrix. We demonstrate that TUM622 acini in 3D coculture recapitulate key features of tissue architecture during LUSC progression as well as the dynamic interactions between LUSC cells and components of the tumor microenvironment (TME), including the extracellular matrix (ECM) and cancer-associated fibroblasts (CAFs). We further adapt our principal 3D culturing protocol to demonstrate how this system could be utilized for various downstream analyses. Overall, this organoid model creates a biologically rich and adaptable platform that enables one to gain insight into the cell-intrinsic and extrinsic mechanisms that promote the disruption of epithelial architectures during carcinoma progression and will aid the search for new therapeutic targets and diagnostic markers.
肿瘤-基质相互作用在肺鳞癌（LUSC）的发展中起着关键作用。然而，由于缺乏合适的模型，了解这些动态相互作用如何导致肿瘤发生过程中观察到的组织结构变化仍然具有挑战性。在本方案中，我们描述了一种使用称为TUM622的LUSC原代细胞培养物生成三维共培养模型的方法。TUM622细胞是从LUSC患者来源的异种移植瘤（PDX）中建立的，当接种在基底膜基质中时具有形成腺泡样结构的独特特性。我们证明，三维共培养中的TUM622腺泡概括了LUSC进展过程中组织结构的关键特征以及LUSC细胞与肿瘤微环境（TME）成分之间的动态相互作用，包括细胞外基质（ECM）和癌症相关成纤维细胞（CAF）。我们进一步调整了主要的三维培养方案，以展示该系统如何用于各种下游分析。总体而言，这种类器官模型创建了一个生物学丰富且适应性强的平台，使人们能够深入了解在癌症进展过程中促进上皮结构破坏的细胞内在和外在机制，并将有助于寻找新的治疗靶点和诊断标志物。
