2D surfaces offer simple analysis of cells in culture, yet these often yield different cell morphologies and responses from those observed in vivo. Considerable effort has therefore been expended on the generation of more tissue-like environments for the study of cell behavior in vitro. Purified matrix proteins provide a 3D scaffold that better mimics the in vivo situation; however, these are far removed from the complex tissue composition seen in vivo. Cell-derived matrices (CDMs) offer a more physiologically relevant alternative for studying in vivo-like cell behavior in vitro. In the protocol described here, fibroblasts cultured on gelatin-coated surfaces are maintained in the presence of ascorbic acid to strengthen matrix deposition over 1-3 weeks. The resulting fibrillar CDMs are denuded of cells, and other cells are subsequently cultured on them, after which their behavior is monitored. We also demonstrate how to use CDMs as an in vivo-relevant reductionist model for studying tumor-stroma-induced changes in carcinoma cell proliferation and migration.

中文翻译：

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在复杂的3D微环境中研究细胞增殖和方向迁移的细胞衍生基质。

2D表面可以对培养中的细胞进行简单分析，但是它们通常会产生与体内观察到的不同的细胞形态和反应。因此，已经花费了相当大的努力来产生更多的类似组织的环境，以用于体外细胞行为的研究。纯化的基质蛋白提供了3D支架，可以更好地模拟体内情况。然而，它们与体内观察到的复杂组织组成相去甚远。细胞衍生基质（CDM）为研究体外类体内细胞行为提供了更生理相关的选择。在此处所述的协议中，将培养在明胶涂层表面上的成纤维细胞在抗坏血酸的存在下进行维护，以在1-3周内增强基质沉积。产生的原纤维CDM被细胞剥夺，然后在其上培养其他细胞，然后监测其行为。我们还演示了如何使用CDM作为体内相关的还原论模型来研究肿瘤基质诱导的癌细胞增殖和迁移的变化。