Analyzing the physicodynamics of immune cells in a three-dimensional collagen matrix.
Analyzing the physicodynamics of immune cells in a three-dimensional collagen matrix.
复制标题
分析三维胶原蛋白基质中免疫细胞的物理动力学。
DOI：
10.1007/978-1-59745-395-0_15
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发表时间：
2007
影响因子：
--
通讯作者：
M. Gunzer
中科院分区：
文献类型：
--
作者：
P. Reichardt;F. Gunzer;M. Gunzer
免疫细胞的运动是其功能不可或缺的先决条件。细胞免疫的所有基本步骤都依赖于细胞迁移和相互作用的能力。虽然在体内观察这些现象将是最生理的方法，但活体成像在技术上要求很高，并且不适合常规或高通量分析。任何良好的体外实验系统都应该反映活组织中细胞迁移和相互作用的内在三维性。在过去十年中产生的数据显示，重要的细胞参数，如细胞速度，细胞形状，和细胞-细胞相互作用的物理动力学非常类似于在三维（3D）胶原蛋白基质测定中测量时在体内观察到的值，其特征在于由I型胶原蛋白组成的水合纤维网络，细胞外基质的主要成分。在这一章中，我们详细描述了3D胶原基质系统的实验使用。我们描述了以小鼠骨髓来源的树突状细胞和抗原特异性辅助性T细胞为例的免疫细胞的制备，3D胶原基质室的建立和使用，细胞迁移和细胞-细胞相互作用的真实的实时荧光显微镜分析的程序，以及由自行开发的计算机辅助细胞跟踪软件支持的数据分析。
The movement of immune cells is an indispensable prerequisite for their function. All essential steps of cellular immunity rely on the ability of cells to migrate and to interact with each other. Although observation of these phenomena in vivo would be the most physiological approach, intravital imaging is technically very demanding and not optimally suited for routine or high-throughput analysis. Any good in vitro experimental system should reflect the inherent three-dimensionality of cell migration and interaction in living tissues. Data generated over the last decade show that important cellular parameters like cell velocity, cell shape, and the physicodynamics of cell-cell interactions closely resemble values observed in vivo when measured in a three-dimensional (3D) collagen matrix assay, featuring a hydrated network of fibers consisting of type I collagen, the major component of the extracellular matrix. In this chapter, we describe in detail the experimental use of the 3D collagen matrix system. We delineate the preparation of immune cells exemplified by bone marrow-derived dendritic cells and antigen specific T-helper cells of the mouse, the build-up and use of the 3D collagen matrix chamber, the procedures of real time fluorescence microscopic analysis of cell migration and cell-cell interaction, as well as data analysis supported by a self-developed software for computer-assisted cell tracking.