Multiplex profiling of cellular invasion in 3D cell culture models.

Multiplex profiling of cellular invasion in 3D cell culture models.
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DOI:
10.1371/journal.pone.0063121
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发表时间:
2013
期刊:
影响因子:
3.7
通讯作者:
Eickelberg O
Eickelberg O
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Burgstaller G;Oehrle B;Koch I;Lindner M;Eickelberg O

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迄今为止,大多数侵入或迁移测定使用改良的Boyden室样设计来评估迁移,作为涂覆或未涂覆的平面塑料表面上的单细胞或划痕测定。在这里,我们描述了一个96孔微孔板为基础的,高含量,三维细胞培养测定能够评估入侵动力学和分子特征。在应用我们的侵袭测定时,我们能够证明对成纤维细胞系以及原代肺成纤维细胞的侵袭能力的显著影响。表皮生长因子的管理导致细胞侵袭的大幅增加,从而使该技术适用于调节原代细胞的侵袭和迁移途径的新型化合物的高通量药理学筛选。我们的测定还将细胞侵袭性与分子事件关联起来。因此,我们认为已经开发了一个强大的和多功能的工具箱,用于在96孔格式中广泛分析侵袭性细胞。这将对纤维化、转移性癌症或慢性炎症状态等疾病领域的研究产生重大影响。
To-date, most invasion or migration assays use a modified Boyden chamber-like design to assess migration as single-cell or scratch assays on coated or uncoated planar plastic surfaces. Here, we describe a 96-well microplate-based, high-content, three-dimensional cell culture assay capable of assessing invasion dynamics and molecular signatures thereof. On applying our invasion assay, we were able to demonstrate significant effects on the invasion capacity of fibroblast cell lines, as well as primary lung fibroblasts. Administration of epidermal growth factor resulted in a substantial increase of cellular invasion, thus making this technique suitable for high-throughput pharmacological screening of novel compounds regulating invasive and migratory pathways of primary cells. Our assay also correlates cellular invasiveness to molecular events. Thus, we argue of having developed a powerful and versatile toolbox for an extensive profiling of invasive cells in a 96-well format. This will have a major impact on research in disease areas like fibrosis, metastatic cancers, or chronic inflammatory states.
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