Spessotto Paola, Giacomello Emiliana, Perri Roberto
Division for Experimental Oncology 2, The National Cancer Institute CRO-IRCCS, Aviano, PN, Italy.
Mol Biotechnol. 2002 Mar;20(3):285-304. doi: 10.1385/mb:20:3:285.
Cell adhesion and cell migration are two primary cellular phenomena to be approached in vitro in order to allow for the effective dissection of the individual events and the unravelling of their underlying molecular mechanisms. The use of assays dedicated to the analysis of cell adhesion and migration in vitro also affords an efficient way of conducting larger basic and applied research screenings of the conditions affecting these processes and are potentially exploitable in the context of routine tests in the biological and medical fields. Therefore, there is a substantial interest in devicing more rationale such assays and major contributions in this direction have been provided by the advent of procedures based on fluorescent cell tagging. In this article we describe three fluorescence-based model assays for the qualitative and quantitative assessment of cell adhesion and cell locomotion in static and dynamic conditions. The assays are easily performed, accurate and reproducible, and can be automatized for high-throughput screenings of cell behavior in vitro. Performance of the assays involves the use of certain dedicated disposable accessories, which are commercially available, and a few instruments that, due to their versatility, can be regarded as constituents of a more generic laboratory setup.
细胞黏附和细胞迁移是两种主要的细胞现象，需要在体外进行研究，以便有效地剖析各个事件并揭示其潜在的分子机制。使用专门用于体外细胞黏附和迁移分析的检测方法，也为对影响这些过程的条件进行更大规模的基础研究和应用研究筛选提供了一种有效的途径，并且在生物和医学领域的常规检测中具有潜在的应用价值。因此，人们对设计更合理的此类检测方法有着浓厚的兴趣，基于荧光细胞标记的方法的出现为此方向做出了重大贡献。在本文中，我们描述了三种基于荧光的模型检测方法，用于在静态和动态条件下对细胞黏附和细胞运动进行定性和定量评估。这些检测方法操作简便、准确且可重复，并且可以自动化以进行体外细胞行为的高通量筛选。检测方法的实施需要使用某些专用的一次性配件，这些配件可在市场上买到，还需要一些仪器，由于其通用性，可以被视为更通用实验室设置的组成部分。
