Mechanobiology Institute, National University of Singapore, Singapore 117411.
EMBL Australia, Single Molecule Science Node, School of Medical Sciences, University of New South Wales 2052, Sydney, Australia.
Mol Biol Cell. 2020 Dec 1;31(25):2779-2790. doi: 10.1091/mbc.E20-07-0466. Epub 2020 Oct 21.
The potential to migrate is one of the most fundamental functions for various epithelial, mesenchymal, and immune cells. Image analysis of motile cell populations, both primary and cultured, typically reveals an intercellular variability in migration speeds. However, cell migration chromatography, the sorting of large populations of cells based on their migratory characteristics, cannot be easily performed. The lack of such methods has hindered our understanding of the direct correlation between the capacity to migrate and other cellular properties. Here, we report two novel, easily implementable and readily scalable methods to sort millions of live migratory cancer and immune cells based on their spontaneous migration in two-dimensional and three-dimensional microenvironments, respectively. Correlative downstream transcriptomic, molecular and functional tests reveal marked differences between the fast and slow subpopulations in patient-derived cancer cells. We further employ our method to reveal that sorting the most migratory cytotoxic T lymphocytes yields a pool of cells with enhanced cytotoxicity against cancer cells. This phenotypic assay opens new avenues for the precise characterization of the mechanisms underlying hither to unexplained heterogeneities in migratory phenotypes within a cell population, and for the targeted enrichment of the most potent migratory leukocytes in immunotherapies.
迁移的潜力是各种上皮细胞、间充质细胞和免疫细胞的最基本功能之一。对运动细胞群体（包括原代和培养的细胞）的图像分析通常揭示了迁移速度的细胞间变异性。然而，基于迁移特性对大量细胞进行分选的细胞迁移色谱法却不容易实现。缺乏这些方法阻碍了我们理解迁移能力与其他细胞特性之间的直接相关性。在这里，我们报告了两种新颖的、易于实施和可扩展的方法，分别基于二维和三维微环境中细胞的自发迁移，对来自癌症和免疫患者的数百万个活迁移细胞进行分选。相关的下游转录组学、分子和功能测试揭示了源自患者的癌症细胞中快速和慢速亚群之间的显著差异。我们进一步利用我们的方法揭示，对最具迁移性的细胞毒性 T 淋巴细胞进行分选，可得到一群对癌细胞具有增强的细胞毒性的细胞。这种表型测定为精确表征迄今为止在细胞群体中迁移表型的未知异质性的机制以及免疫治疗中最有效的迁移白细胞的靶向富集开辟了新的途径。
