Nanobioengineering Group, Institute for Bioengineering of Catalonia (IBEC) , Baldiri Reixac 15-21, 08028 Barcelona, Spain.
Department of Engineering: Electronics, University of Barcelona , 08028 Barcelona, Spain.
ACS Appl Mater Interfaces. 2017 Feb 1;9(4):4159-4170. doi: 10.1021/acsami.6b13063. Epub 2017 Jan 18.
The interaction of cells with their natural environment influences a large variety of cellular phenomena, including cell adhesion, proliferation, and migration. The complex extracellular matrix network has challenged the attempts to replicate in vitro the heterogeneity of the cell environment and has threatened, in general, the relevance of in vitro studies. In this work, we describe a new and extremely versatile approach to generate native-like extracellular matrices with controlled morphologies for the in vitro study of cellular processes. This general approach combines the confluent culture of fibroblasts with microfabricated guiding templates to direct the three-dimensional growth of well-defined extracellular networks which recapitulate the structural and biomolecular complexity of features typically found in vivo. To evaluate its performance, we studied fundamental cellular processes, including cell cytoskeleton organization, cell-matrix adhesion, proliferation, and protrusions morphodynamics. In all cases, we found striking differences depending on matrix architecture and, in particular, when compared to standard two-dimensional environments. We also assessed whether the engineered matrix networks influenced cell migration dynamics and locomotion strategy, finding enhanced migration efficiency for cells seeded on aligned matrices. Altogether, our methodology paves the way to the development of high-performance models of the extracellular matrix for potential applications in tissue engineering, diagnosis, or stem-cell biology.
细胞与其自然环境的相互作用影响着多种细胞现象，包括细胞黏附、增殖和迁移。复杂的细胞外基质网络给试图在体外复制细胞环境的异质性带来了挑战，并威胁到体外研究的相关性。在这项工作中，我们描述了一种新的、非常通用的方法，可以生成具有控制形态的天然样细胞外基质，用于体外研究细胞过程。这种通用方法结合了成纤维细胞的致密培养和微加工导向模板，以指导三维生长出具有明确细胞外网络，这些网络再现了体内通常发现的结构和生物分子复杂性特征。为了评估其性能，我们研究了基本的细胞过程，包括细胞细胞骨架组织、细胞基质黏附、增殖和突起形态动力学。在所有情况下，我们发现根据基质结构会有显著差异，特别是与标准二维环境相比时。我们还评估了工程化的基质网络是否会影响细胞迁移动力学和运动策略，发现细胞在排列的基质上的迁移效率更高。总的来说，我们的方法为细胞外基质的高性能模型的开发铺平了道路，这些模型可能在组织工程、诊断或干细胞生物学中有潜在应用。
