High-Content Analysis of Cell Migration Dynamics within a Micropatterned Screening Platform.
High-Content Analysis of Cell Migration Dynamics within a Micropatterned Screening Platform.
复制标题
微图案筛选平台内细胞迁移动力学的高内涵分析。
DOI：
10.1002/adbi.201900011
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发表时间：
2019
影响因子：
4.1
通讯作者：
Almeida FV
中科院分区：
文献类型：
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作者：
Almeida FV
细胞迁移是一个基本的生物学过程，它受到微环境和内在基因表达程序之间复杂相互作用的动态调节。在这里，使用微图案化和动态粘附的聚合物刷基底开发了高通量细胞迁移测定，其支持对96孔细胞培养板格式内的细胞-基质相互作用的高度精确和一致的控制。该系统与细胞运动性和F-肌动蛋白细胞骨架组织的自动成像和定量相结合，用于细胞迁移表型的高含量分析。使用该平台筛选147种表观遗传抑制剂的文库，鉴定出一组EZH 2特异性化合物，其通过上皮向间充质转化样基因表达程序的去抑制来促进细胞骨架重塑并加速角质形成细胞迁移。总之，这些研究建立了高通量，微模式化测定作为一个强大的工具，用于发现新的治疗靶点和解剖复杂的基因-环境相互作用参与伤口修复。
Cell migration is a fundamental biological process that is dynamically regulated by complex interactions between the microenvironment and intrinsic gene expression programs. Here, a high‐throughput cell migration assay is developed using micropatterned and dynamically adhesive polymer brush substrates, which support highly precise and consistent control over cell–matrix interactions within a 96‐well cell culture plate format. This system is combined with automated imaging and quantitation of both cell motility and organization of the F‐actin cytoskeleton for high‐content analysis of cell migration phenotypes. Using this platform to screen a library of 147 epigenetic inhibitors identifies a set of EZH2‐specific compounds that promote cytoskeletal remodeling and accelerates keratinocyte migration through derepression of an epithelial to mesenchymal transition‐like gene expression program. Together, these studies establish the high‐throughput, micropatterned assay as a powerful tool for discovery of novel therapeutic targets and for dissecting complex gene–environment interactions involved in wound repair.