Costa Erico Tosoni, Camargo Anamaria Aranha
Centro de Oncologia Molecular, Hospital Sírio Libanês, São Paulo, Brazil.
Methods Mol Biol. 2018;1731:155-168. doi: 10.1007/978-1-4939-7595-2_15.
In this chapter, we describe a straightforward protocol to generate multicellular tumor spheroids (MTSs) and evaluate the role of specific genes in regulating cell invasiveness in real-time and tridimensional (3D) matrices. This approach provides advantages over other conventional invasion assays by offering intimate cell-cell and cell-ECM contacts and by mimicking the pathophysiological characteristics observed in tumor microenvironments (e.g., microregional gradients in glucose and O concentrations and metabolic and proliferative tumor heterogeneity). We also provide an original and semiautomated approach to quantify MTS invasion using the freely available ImageJ software and plugins.
在本章中，我们描述了一种简单的方案，用于生成多细胞肿瘤球体（MTSs），并实时评估特定基因在三维（3D）基质中调节细胞侵袭性的作用。与其他传统侵袭试验相比，这种方法具有优势，它提供了紧密的细胞间和细胞与细胞外基质（ECM）的接触，并模拟了肿瘤微环境中观察到的病理生理特征（例如，葡萄糖和氧气浓度的微区域梯度以及肿瘤的代谢和增殖异质性）。我们还提供了一种原始的半自动化方法，使用免费的ImageJ软件和插件来量化MTS的侵袭。
