Department of Biomedical Engineering, Gachon University, Seongnam 13120, Korea.
Cytek Biosciences, 47215 Lakeview Blvd, Fremont, CA 95348, USA.
Molecules. 2022 Aug 19;27(16):5306. doi: 10.3390/molecules27165306.
The 3D cell migration assay was developed for the evaluation of drugs that inhibit cell migration using high throughput methods. Wound-healing assays have commonly been used for cell migration assays. However, these assays have limitations in mimicking the in vivo microenvironment of the tumor and measuring cell viability for evaluation of cell migration inhibition without cell toxicity. As an attempt to manage these limitations, cells were encapsulated with Matrigel on the surface of the pillar, and an analysis of the morphology of cells attached to the pillar through Matrigel was performed for the measurement of cell migration. The micropillar/microwell chips contained 532 pillars and wells, which measure the migration and viability of cells by analyzing the roundness and size of the cells, respectively. Cells seeded in Matrigel have a spherical form. Over time, cells migrate through the Matrigel and attach to the surface of the pillar. Cells that have migrated and adhered have a diffused shape that is different from the initial spherical shape. Based on our analysis of the roundness of the cells, we were able to distinguish between the diffuse and spherical shapes. Cells in Matrigel on the pillar that were treated with migration-inhibiting drugs did not move to the surface of the pillar and remained in spherical forms. During the conduct of experiments, 70 drugs were tested in single chips and migration-inhibiting drugs without cell toxicity were identified. Conventional migration assays were performed using transwell for verification of the four main migration-inhibiting drugs found on the chip.
三维细胞迁移分析用于评估使用高通量方法抑制细胞迁移的药物。划痕实验常用于细胞迁移分析。然而,这些实验在模拟肿瘤体内微环境和测量细胞活力以评估细胞迁移抑制而不产生细胞毒性方面存在局限性。作为对这些局限性的尝试性管理,细胞用 Matrigel 包被在柱子表面,通过分析附着在柱子上的细胞的形态来测量细胞迁移。微柱/微孔芯片包含 532 个柱子和孔,通过分析细胞的圆度和大小分别测量细胞的迁移和活力。在 Matrigel 中接种的细胞呈球形。随着时间的推移,细胞穿过 Matrigel 并附着在柱子表面。迁移并附着的细胞具有与初始球形不同的弥散形状。基于我们对细胞圆度的分析,我们能够区分弥散形状和球形。用迁移抑制药物处理的柱子上的 Matrigel 中的细胞不会移动到柱子表面,而是保持球形。在实验过程中,在单个芯片上测试了 70 种药物,并确定了没有细胞毒性的迁移抑制药物。使用 Transwell 进行常规迁移实验,以验证芯片上发现的四种主要迁移抑制药物。