Preparation of 3D Collagen Gels and Microchannels for the Study of 3D Interactions In Vivo
Preparation of 3D Collagen Gels and Microchannels for the Study of 3D Interactions In Vivo
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DOI：
10.3791/53989
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发表时间：
2016-05-01
影响因子：
1.2
通讯作者：
Keely, Patricia J.
中科院分区：
文献类型：
--
作者：
Burkel, Brian;Morris, Brett A.;Keely, Patricia J.
从历史上看，大多数细胞过程仅在2个维度上进行了研究。尽管这些研究对通用细胞信号传导机制具有丰富的信息，但它们忽略了从局部微环境和细胞外基质（ECM）的结构和机械性能中获得的重要细胞提示。为了了解细胞在生理ECM中的相互作用，在3个维测定的背景下研究它们很重要。细胞迁移，细胞分化和细胞增殖只是3维ECM机械性能的局部变化所影响的几个过程。 ECM的核心原纤维成分胶原I不仅仅是组织的简单结构元素。在正常条件下，胶原网络直接形态发生并维持细胞结构的机械提示。在患病的微环境（例如肿瘤微环境）中，胶原蛋白网络通常经常重塑，表明组成改变，增强的沉积和纤维组织改变。在乳腺癌中，纤维比对的程度很重要，因为垂直于肿瘤边界的对齐纤维的增加与患者的预后较差有关（1）。对齐的胶原蛋白矩阵导致通过持续迁移增加肿瘤细胞的传播（2,3）。以下是将细胞嵌入三维纤维胶原水凝胶中的简单方案。该方案很容易适应许多平台，并且可以在可调的，三维的生理微环境中生成对齐和随机胶原蛋白矩阵，以研究细胞迁移，细胞分裂和其他细胞过程。
Historically, most cellular processes have been studied in only 2 dimensions. While these studies have been informative about general cell signaling mechanisms, they neglect important cellular cues received from the structural and mechanical properties of the local microenvironment and extracellular matrix (ECM). To understand how cells interact within a physiological ECM, it is important to study them in the context of 3 dimensional assays. Cell migration, cell differentiation, and cell proliferation are only a few processes that have been shown to be impacted by local changes in the mechanical properties of a 3-dimensional ECM. Collagen I, a core fibrillar component of the ECM, is more than a simple structural element of a tissue. Under normal conditions, mechanical cues from the collagen network direct morphogenesis and maintain cellular structures. In diseased microenvironments, such as the tumor microenvironment, the collagen network is often dramatically remodeled, demonstrating altered composition, enhanced deposition and altered fiber organization. In breast cancer, the degree of fiber alignment is important, as an increase in aligned fibers perpendicular to the tumor boundary has been correlated to poorer patient prognosis(1). Aligned collagen matrices result in increased dissemination of tumor cells via persistent migration(2,3). The following is a simple protocol for embedding cells within a 3-dimensional, fibrillar collagen hydrogel. This protocol is readily adaptable to many platforms, and can reproducibly generate both aligned and random collagen matrices for investigation of cell migration, cell division, and other cellular processes in a tunable, 3-dimensional, physiological microenvironment.