Norwegian University of Science and Technology (NTNU), Department of Clinical and Molecular Medicine, NO-7491 Trondheim, Norway.
Norwegian University of Science and Technology (NTNU), Department of Clinical and Molecular Medicine, NO-7491 Trondheim, Norway; Korsnes Biocomputing (KoBio), Trondheim, Norway.
Exp Cell Res. 2024 Apr 1;437(1):113993. doi: 10.1016/j.yexcr.2024.113993. Epub 2024 Mar 13.
This article demonstrates that label-free single-cell video tracking is a useful approach for in vitro studies of Epithelial-Mesenchymal Transition (EMT). EMT is a highly heterogeneous process, involved in wound healing, embryogenesis and cancer. The process promotes metastasis, and increased understanding can aid development of novel therapeutic strategies. The role of EMT-associated biomarkers depends on biological context, making it challenging to compare and interpret data from different studies. We demonstrate single-cell video tracking for comprehensive phenotype analysis. In this study we performed single-cell video tracking on 72-h long recordings. We quantified several behaviours at a single-cell level during induced EMT in MDA-MB-468 cells. This revealed notable variations in migration speed, with different dose-response patterns and varying distributions of speed. By registering cell morphologies during the recording, we determined preferred paths of morphological transitions. We also found a clear association between migration speed and cell morphology. We found elevated rates of cell death, diminished proliferation, and an increase in mitotic failures followed by re-fusion of sister-cells. The method allows tracking of phenotypes in cell lineages, which can be particularly useful in epigenetic studies. Sister-cells were found to have significant similarities in their speeds and morphologies, illustrating the heritability of these traits.
本文证明，无标记的单细胞视频跟踪是体外研究上皮-间充质转化 (EMT) 的有用方法。EMT 是一个高度异质的过程，涉及伤口愈合、胚胎发生和癌症。该过程促进转移，增加对其的理解有助于开发新的治疗策略。EMT 相关生物标志物的作用取决于生物学背景，因此比较和解释来自不同研究的数据具有挑战性。我们展示了用于全面表型分析的单细胞视频跟踪。在这项研究中，我们对 72 小时长的记录进行了单细胞视频跟踪。我们在 MDA-MB-468 细胞中诱导 EMT 期间在单细胞水平上量化了几种行为。这揭示了迁移速度的显著变化，具有不同的剂量反应模式和不同的速度分布。通过在记录过程中注册细胞形态，我们确定了形态转变的首选路径。我们还发现迁移速度和细胞形态之间存在明显的关联。我们发现细胞死亡率升高、增殖减少以及有丝分裂失败增加，随后姐妹细胞重新融合。该方法允许在细胞谱系中跟踪表型，这在表观遗传学研究中特别有用。发现姐妹细胞在速度和形态上具有显著的相似性，说明这些特征具有遗传性。
