Neutrophil Migration Through Extracellular Matrix
Neutrophil Migration Through Extracellular Matrix
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DOI：
10.1007/978-1-62703-845-4_13
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发表时间：
2014-01-01
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影响因子：
--
通讯作者：
Knaus, Ulla G.
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文献类型：
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作者：
Jennings, Richard T.;Knaus, Ulla G.
神经元从血流迁移到感染或损伤部位是一个多步骤的过程，取决于所遇到的组织结构，需要不同的运动模式。神经元运动的范围可以从间充质到变形虫运动，并且可以包括多种形状变化、通过间隙的收缩挤压以及粘附/去粘附循环。体外迁移试验仅反映了复杂的体内中性粒细胞募集的某些方面。对于二维体外迁移趋化性室，使用移液器梯度或Boyden室移动的显微镜分析。为了分析三维体外迁移，可以将中性粒细胞包埋到具有不同生物物理特性的基质中，或者可以将其置于在宽孔过滤器上分层的基质上，使得能够朝向化学引诱物的梯度迁移通过transwell板的基质和过滤器。我们在此利用市售的装置，用于从松散的I型胶原基质的顶部迁移鼠嗜中性粒细胞，其确定嗜中性粒细胞附着于基质、感测化学引诱物、消化、消化基质并通过基质移动到下部transwell室的能力。虽然不能详细研究基质内的迁移模式，但该试验允许在规定的时间段内定量评估迁移的中性粒细胞。
Neutrophil migration from the bloodstream to sites of infection or injury is a multistep process that requires, dependent on the tissue structures being encountered, different modes of movement. Neutrophil locomotion can range from mesenchymal to amoeboid movement and may include multiple shape changes, contractile squeezing through gaps, and adhesion/de-adhesion cycles. In vitro migration assays reflect only some aspects of the complex in vivo neutrophil recruitment. For two-dimensional in vitro migration chemotaxis chambers, microscopic analysis of movement towards a pipette gradient or Boyden chambers is used. To analyze three-dimensional in vitro migration neutrophils can be embedded into matrices of diverse biophysical properties or can be placed onto matrices that are layered on a wide-pore filter, enabling migration through the matrix and the filter of a transwell plate towards a gradient of chemoattractant. We utilize here a commercially available setup for migration of murine neutrophils from the top of a loose collagen type I matrix, which determines the ability of neutrophils to attach to the matrix, sense the chemoattractant, polarize, digest the matrix, and move through the matrix into the lower transwell chamber. While the mode of migration inside the matrix cannot be studied in detail, this assay permits quantitative assessment of migrated neutrophils during a defined period of time.