Visweshwaran Sai P, Gautreau Alexis
Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle-Straße 10, Berlin, Germany.
BIOC, CNRS UMR7654, Ecole Polytechnique, Institut Polytechnique de Paris, Palaiseau, France.
Bio Protoc. 2020 Jan 5;10(1):e3482. doi: 10.21769/BioProtoc.3482.
The ability of cancer cells to migrate through a complex three-dimensional (3D) environment is a hallmark event of cancer metastasis. Therefore, an migration assay to evaluate cancer cell migration in a 3D setting is valuable to examine cancer progression. Here, we describe such a simple migration assay in a 3D collagen-fibronectin gel for observing cell morphology and comparing the migration abilities of cancer cells. We describe below how to prepare the collagen-fibronectin gel castings, how to set up time-lapse recording, how to draw single-cell trajectories from movies and extract key parameters that characterize cell motility, such as cell speed, directionality, mean square displacement, and directional persistence. In our set-up, cells are sandwiched in a single plane between two collagen-fibronectin gels. This trick facilitates the analysis of cell tracks, which are for the most part 2D, at least in the beginning, but in a 3D environment. This protocol has been previously published in Visweshwaran (2018) and is described here in more detail.
癌细胞在复杂的三维（3D）环境中迁移的能力是癌症转移的标志性事件。因此，一种用于评估癌细胞在三维环境中迁移的迁移实验，对于研究癌症进展具有重要价值。在此，我们描述一种在三维胶原蛋白-纤连蛋白凝胶中进行的简单迁移实验，用于观察细胞形态并比较癌细胞的迁移能力。下面我们将介绍如何制备胶原蛋白-纤连蛋白凝胶铸型，如何设置延时记录，如何从视频中绘制单细胞轨迹以及提取表征细胞运动性的关键参数，如细胞速度、方向性、均方位移和方向持续性。在我们的实验设置中，细胞夹在两个胶原蛋白-纤连蛋白凝胶之间的单个平面中。这个技巧便于分析细胞轨迹，这些轨迹在很大程度上是二维的，至少在开始时是这样，但处于三维环境中。该方案先前已发表于维斯韦什瓦兰（2018年），此处将更详细地进行描述。
