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Generation of 3D Tumor Spheroids with Encapsulating Basement Membranes for Invasion Studies

机译：用封装基底膜的3D肿瘤球体产生侵袭性研究

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In the past, in vitro studies of invasion and tumor progression were performed primarily using cancer cells cultured on a flat, two‐dimensional (2D) surface in a monolayer. In recent years, however, many studies have demonstrated differences in cellsignaling and cell migration between 2D and 3D cell cultures. Traditional 2D monolayer cancer cell invasion models do not fully recapitulate 3D cell‐to‐cell and cell-to-extracellular matrix interactions that in vivo models can provide. Moreover, although in vivo animal models are irreplaceable for studying tumor biology and metastasis, they are costly, time‐consuming, and impractical for answering preliminary questions. Thus, emergent and evolving 3D spheroid cell culture models have changed the waywe study tumors and their interactions with their surrounding extracellular matrix. In the case of breast cancer, metastasis of breast cancer tumors results in high mortality rates, and thus development of robust cell culture models that are reproducibleand practical for studying breast cancer progression is important for ultimately developing preventatives for cancer metastasis. This article provides a set of protocols for generating uniform spheroids with a thin sheet of basement membrane for studying the initial invasion of mammary epithelial cells into a surrounding collagen‐rich extracellular matrix. Details are provided for generating 3D spheroids with a basement membrane, polymerizing collagen I, embedding the spheroids in the 3D collagen gel,and immunostaining the spheroids for invasion studies.
机译：在过去，主要使用在单层中的平坦的二维（2D）表面上培养的癌细胞进行侵袭和肿瘤进展的体外研究。然而，近年来，许多研究表明，2D和3D细胞培养物之间的细胞系列和细胞迁移差异。传统的2D单层癌细胞侵袭模型不完全重新承载3D细胞到细胞和细胞对细胞的基质相互作用，其在体内模型可以提供。此外，尽管在体内动物模型中是不可替代的，但对于研究肿瘤生物学和转移，但它们是昂贵，耗时的，并且对于回答初步问题而言是不切实际的。因此，紧急和不断发展的3D球形细胞培养模型改变了研究肿瘤和与周围细胞外基质的相互作用。在乳腺癌的情况下，乳腺癌肿瘤的转移导致高死亡率，因此对研究乳腺癌进展的再生能力的强大细胞培养模型的发展对于最终产生癌症转移的预防性是重要的。本文提供了一组用于产生均匀球体的协议，其具有薄的基底膜，用于研究乳腺上皮细胞的初始侵袭到富含胶原蛋白的细胞外基质中。提供细节用于产生具有基底膜的3D球状体，聚合胶原I，将球状体嵌入3D胶原凝胶中，并免疫染色球体用于侵入研究。

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