Shabalina Evgenya Y, Skorova Ekaterina Yu, Chudakova D A, Anikin V B, Reshetov I V, Mynbaev O A, Petersen E V
Moscow Institute of Physics and Technology, Institutskiy Pereulok, Dolgoprudny, Russia.
School of Biological Sciences, University of Auckland, Auckland, New Zealand.
Front Mol Biosci. 2021 Mar 25;8:610407. doi: 10.3389/fmolb.2021.610407. eCollection 2021.
3D cell culture systems utilizing multicellular tumor spheroids (MCTS) are widely used in translational oncology, including for studying cell migration and in personalized therapy. However, early stages of cellular migration from MCTS and cross-talk between spheroids are overlooked, which was addressed in the current study. Here, we investigated cell migration from MCTS derived from human non-small cell lung cancer (NSCLC) cell line A549 cultured on different substrates, collagen gel or plastic, at different time points. We found that migration starts at 4-16 h time points after the seeding and its speed is substrate-dependent. We also demonstrated that co-culture of two NSCLC-derived MCTS on collagen gel, but not on plastic, facilitates cell migration compared with single MTCS. This finding should be considered when designing MCTS-based functional assays for personalized therapeutic approach and drug screenings. Overall, our work characterizes the 3D cell culture model resembling NSCLC cell migration from the clusters of CTCs into surgical wound, and describes microscopy-based tools and approaches for image data analysis with a potential for further automation. These tools and approaches also might be used to predict patterns of CTCs migration based on analysis of patient biopsy in a 3D culture system.
利用多细胞肿瘤球体（MCTS）的3D细胞培养系统在转化肿瘤学中被广泛应用，包括用于研究细胞迁移和个性化治疗。然而，MCTS细胞迁移的早期阶段以及球体之间的相互作用被忽视了，本研究解决了这一问题。在这里，我们研究了在不同时间点，源自人非小细胞肺癌（NSCLC）细胞系A549且培养在不同底物（胶原凝胶或塑料）上的MCTS的细胞迁移情况。我们发现迁移在接种后4 - 16小时的时间点开始，其速度取决于底物。我们还证明，与单个MCTS相比，在胶原凝胶上而非塑料上共同培养两个源自NSCLC的MCTS可促进细胞迁移。在设计基于MCTS的个性化治疗方法和药物筛选功能测定时应考虑这一发现。总体而言，我们的工作描述了类似于NSCLC细胞从循环肿瘤细胞（CTC）簇迁移到手术伤口的3D细胞培养模型，并描述了基于显微镜的图像数据分析工具和方法，具有进一步自动化的潜力。这些工具和方法也可用于基于3D培养系统中患者活检分析来预测CTC迁移模式。
