CAFs and Cancer Cells Co-Migration in 3D Spheroid Invasion Assay
CAFs and Cancer Cells Co-Migration in 3D Spheroid Invasion Assay
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DOI：
10.1007/978-1-0716-0779-4_19
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发表时间：
2021-01-01
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影响因子：
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通讯作者：
Labernadie, Anna
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文献类型：
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作者：
Conti, Sefora;Kato, Takuya;Labernadie, Anna
在许多实体肿瘤中，集体细胞侵袭优于单细胞播散策略。集体侵袭模式通常显示特定的前/后细胞组织，其中侵袭性领导细胞来自癌细胞群或肿瘤基质。集体侵袭涉及协调的细胞运动，需要通过细胞-细胞相互作用和细胞-基质粘附的特定组合进行紧密的机械串扰。癌症相关成纤维细胞（CAFs）最近被报道通过ECM重塑和直接细胞间接触驱动上皮癌细胞的传播。然而，肿瘤和间质细胞之间的相互作用仍然知之甚少。在这里，我们提出了一种简单的球体侵袭试验来评估CAFs在上皮肿瘤细胞集体迁移中的作用。该方法可以通过活细胞荧光标记结合旋转圆盘显微镜来表征三维球体入侵模式。当嵌入细胞外基质时，可以跟踪球体的侵入链，并可以量化CAFs和癌细胞的领导者/追随者组织。
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.