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 2D and 3D 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 hitherto unexplained heterogeneities in migratory phenotypes within a cell population, and for the targeted enrichment of the most potent migratory leukocytes in immunotherapies.

迁移潜力是各种上皮细胞、间充质细胞和免疫细胞最基本的功能之一。原代和培养的运动细胞群的图像分析通常揭示迁移速度的细胞间变异。然而，细胞迁移色谱法（根据细胞迁移特征对大量细胞进行分类）并不容易进行。缺乏此类方法阻碍了我们对迁移能力与其他细胞特性之间直接相关性的理解。在这里，我们报告了两种新颖、易于实施和易于扩展的方法，分别根据数百万活迁移性癌症和免疫细胞在 2D 和 3D 微环境中的自发迁移进行分类。相关的下游转录组、分子和功能测试揭示了患者来源的癌细胞中快亚群和慢亚群之间的显着差异。我们进一步利用我们的方法揭示了分选最具迁移性的细胞毒性 T 淋巴细胞会产生一组对癌细胞具有增强细胞毒性的细胞。这种表型测定为精确表征细胞群内迁移表型中迄今为止无法解释的异质性的机制以及免疫疗法中最有效的迁移白细胞的靶向富集开辟了新途径。