Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute for Science and Technology (BIST), Barcelona, Spain.
The Francis Crick Institute, London, UK.
Methods Mol Biol. 2021;2179:243-256. doi: 10.1007/978-1-0716-0779-4_19.
In many solid tumors, collective cell invasion prevails over single-cell dissemination strategies. Collective modes of invasion often display specific front/rear cellular organization, where invasive leader cells arise from cancer cell populations or the tumor stroma. Collective invasion involves coordinated cellular movements which require tight mechanical crosstalk through specific combinations of cell-cell interactions and cell-matrix adhesions. Cancer Associated Fibroblasts (CAFs) have been recently reported to drive the dissemination of epithelial cancer cells through ECM remodeling and direct intercellular contact. However, the cooperation between tumor and stromal cells remains poorly understood. Here we present a simple spheroid invasion assay to assess the role of CAFs in the collective migration of epithelial tumor cells. This method enables the characterization of 3D spheroid invasion patterns through live cell fluorescent labeling combined with spinning disc microscopy. When embedded in extracellular matrix, the invasive strands of spheroids can be tracked and leader/follower organization of CAFs and cancer cells can be quantified.
在许多实体肿瘤中，细胞的集体浸润胜过单细胞扩散策略。集体浸润模式通常表现出特定的前后细胞组织，其中浸润的主导细胞来源于癌细胞群体或肿瘤基质。集体浸润涉及协调的细胞运动，这需要通过特定的细胞-细胞相互作用和细胞-基质黏附组合来实现紧密的机械串扰。最近有报道称，癌症相关成纤维细胞（CAFs）通过 ECM 重塑和直接细胞间接触来驱动上皮癌细胞的扩散。然而，肿瘤细胞和基质细胞之间的合作仍知之甚少。在这里，我们提出了一种简单的球体浸润测定法来评估 CAFs 在上皮肿瘤细胞集体迁移中的作用。这种方法可以通过活细胞荧光标记结合旋转圆盘显微镜来描述 3D 球体浸润模式。当嵌入细胞外基质时，可以跟踪球体浸润的丝状结构，并定量 CAFs 和癌细胞的主导/跟随组织。
