Cell dissemination during tumor development is a characteristic of cancer metastasis. Dissemination from three-dimensional spheroid models on extracellular matrices designed to mimic tissue-specific physiological microenvironments may allow us to better elucidate the mechanism behind cancer metastasis and the response to therapeutic agents. The orientation of fibrillar collagen plays a key role in cellular processes and mediates metastasis through contact-guidance. Understanding how cells migrate on aligned collagen fibrils requires in vitro assays with reproducible and standardized orientation of collagen fibrils on the macro-to-nanoscale. Herein, we implement a spheroid-based migration assay, integrated with a fibrillar type I collagen matrix, in a manner compatible with high throughput image acquisition and quantitative analysis. The migration of highly proliferating U2OS osteosarcoma cell spheroids onto an aligned fibrillar type I collagen matrix was quantified. Cell dissemination from the spheroid was polarized with increased invasion in the direction of fibril alignment. The resulting area of cell dissemination had an aspect ratio of 1.2 ± 0.1 and an angle of maximum invasion distance of 5° ± 44° relative to the direction of collagen fibril alignment. The assay described here can be applied to a fully automated imaging and analysis pipeline for the assessment of tumor cell migration with high throughput screening.

肿瘤发展过程中的细胞播散是癌症转移的一个特征。在旨在模拟组织特异性生理微环境的细胞外基质上传播三维球体模型可能使我们能够更好地阐明癌症转移背后的机制和对治疗剂的反应。纤维状胶原的取向在细胞过程中起关键作用，并通过接触引导介导转移。了解细胞如何在对齐的胶原原纤维上迁移需要体外在宏观到纳米尺度上具有可重复和标准化方向的胶原原纤维的测定。在这里，我们以与高通量图像采集和定量分析兼容的方式实施了一种基于球体的迁移测定，与纤维状 I 型胶原基质相结合。量化了高度增殖的 U2OS 骨肉瘤细胞球体迁移到对齐的纤维状 I 型胶原基质上。来自球体的细胞传播随着原纤维排列方向的侵入增加而极化。由此产生的细胞传播区域的纵横比为 1.2 ± 0.1，相对于胶原原纤维排列方向的最大侵入距离角度为 5° ± 44°。