Almici Enrico, Caballero David, Montero Boronat Joan, Samitier Martí Josep
Nanobioengineering Group, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), Barcelona, Spain; Department of Electronics and Biomedical Engineering, University of Barcelona, Barcelona, Spain; Centro de Investigación Biomédica en Red en Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Madrid, Spain.
Nanobioengineering Group, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), Barcelona, Spain; Department of Electronics and Biomedical Engineering, University of Barcelona, Barcelona, Spain; Centro de Investigación Biomédica en Red en Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Madrid, Spain.
Methods Cell Biol. 2020;156:161-183. doi: 10.1016/bs.mcb.2019.11.022. Epub 2020 Jan 20.
The composition and architecture of the extracellular matrix (ECM) and their dynamic alterations, play an important regulatory role on numerous cellular processes. Cells embedded in 3D scaffolds show phenotypes and morphodynamics reminiscent of the native scenario. This is in contrast to flat environments, where cells display artificial phenotypes. The structural and biomolecular properties of the ECM are critical in regulating cell behavior via mechanical, chemical and topological cues, which induce cytoskeleton rearrangement and gene expression. Indeed, distinct ECM architectures are encountered in the native stroma, which depend on tissue type and function. For instance, anisotropic geometries are associated with ECM degradation and remodeling during tumor progression, favoring tumor cell invasion. Overall, the development of innovative in vitro ECM models of the ECM that reproduce the structural and physicochemical properties of the native scenario is of upmost importance to investigate the mechanistic determinants of tumor dissemination. In this chapter, we describe an extremely versatile technique to engineer three-dimensional (3D) matrices with controlled architectures for the study of pathophysiological processes in vitro. To this aim, a confluent culture of "sacrificial" fibroblasts was seeded on top of microfabricated guiding templates to induce the 3D ECM growth with specific isotropic or anisotropic architectures. The resulting matrices, and cells seeded on them, recapitulated the structure, composition, phenotypes and morphodynamics typically found in the native scenario. Overall, this method paves the way for the development of in vitro ECMs for pathophysiological studies with potential clinical relevance.
细胞外基质（ECM）的组成、结构及其动态变化，对众多细胞过程发挥着重要的调节作用。嵌入三维支架中的细胞表现出的表型和形态动力学让人联想到天然环境。这与扁平环境形成对比，在扁平环境中细胞呈现出人工表型。ECM的结构和生物分子特性通过机械、化学和拓扑线索调节细胞行为，这些线索会诱导细胞骨架重排和基因表达。事实上，在天然基质中会遇到不同的ECM结构，这取决于组织类型和功能。例如，各向异性的几何结构与肿瘤进展过程中的ECM降解和重塑相关，有利于肿瘤细胞侵袭。总体而言，开发能够重现天然环境结构和物理化学特性的创新型体外ECM模型对于研究肿瘤扩散的机制决定因素至关重要。在本章中，我们描述了一种极其通用的技术，用于构建具有可控结构的三维（3D）基质，以研究体外病理生理过程。为此，将汇合培养的“牺牲性”成纤维细胞接种在微制造的引导模板上，以诱导具有特定各向同性或各向异性结构的3D ECM生长。所得的基质以及接种在其上的细胞，重现了天然环境中常见的结构、组成、表型和形态动力学。总体而言，该方法为开发具有潜在临床相关性的用于病理生理研究的体外ECM铺平了道路。
