Knöll Bernd, Weinl Christine, Nordheim Alfred, Bonhoeffer Friedrich
Interfaculty Institute for Cell Biology, Department of Molecular Biology, University of Tübingen, Auf der Morgenstelle 15, 72076 Tübingen, Germany.
Nat Protoc. 2007;2(5):1216-24. doi: 10.1038/nprot.2007.157.
Stripe assays have been widely employed as in vitro test systems to study the responses of growing axons, as well as migrating cells, to established or novel guidance molecules. We provide detailed protocols for both the original and the modified version of this assay, as they allow the analysis of the 'guidance properties' of active components present in crude membrane fractions or as purified molecules. Silicon matrices are used to produce striped patterns of active molecules on a surface (referred to as 'carpet'), followed by culturing of neurons, or any other cell type, on these carpets. After 1-2 days in culture, striped outgrowth of extending neurites--indicative of guided migration of cell processes--can be observed. We also discuss potential other applications (e.g., in neuronal regeneration and development) and modifications of the assay. The preparation of 10-12 carpets takes approximately 4-5 h.
条纹分析已被广泛用作体外测试系统,以研究生长中的轴突以及迁移细胞对既定或新型导向分子的反应。我们提供了该分析原始版本和修改版本的详细方案,因为它们能够分析粗膜组分中存在的活性成分或纯化分子的“导向特性”。使用硅基质在表面(称为“地毯”)上产生活性分子的条纹图案,然后在这些“地毯”上培养神经元或任何其他细胞类型。培养1-2天后,可以观察到延伸的神经突出现条纹状生长,这表明细胞突起发生了导向迁移。我们还讨论了该分析的其他潜在应用(例如,在神经元再生和发育方面)以及修改方法。制备10-12个“地毯”大约需要4-5小时。