Preparation of High-Density Fibrillar Collagen Matrices That Mimic Desmoplastic Tumor Stroma.
Preparation of High-Density Fibrillar Collagen Matrices That Mimic Desmoplastic Tumor Stroma.
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DOI：
10.1002/0471143030.cb1019s70
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发表时间：
2016-03-01
影响因子：
--
通讯作者：
Artym VV
中科院分区：
文献类型：
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作者：
Artym VV
由于促纤维增生反应，浸润性肿瘤的间质变得富含致密的纤维状胶原。这种促结缔组织增生的胶原蛋白对肿瘤和正常细胞产生深远的影响。鉴于这些发现，重要的是开发新的体外细胞系统，模拟这种促纤维增生细胞外基质，以允许在体内样条件下的细胞研究。本单元提供了一个协议和故障排除指南的准备高密度纤维胶原（HDFC）矩阵，密切模拟结缔组织胶原基质的恶性肿瘤。然后，它描述了使用这种HDFC矩阵在体外细胞研究的侵袭伪足的形成和功能的细胞外基质的入侵。此外，该单位还提供了一个详细的协议，免疫标记的invadopodial蛋白和检测HDFC基质降解与invadopodia允许invadopodia可视化使用荧光显微镜。
The stroma of invasive tumors becomes enriched in dense fibrillar collagen as a result of the desmoplastic reaction. This desmoplastic collagen exerts profound effects on tumor and normal cells. In view of these findings, it is important to develop novel in vitro cell systems that mimic this desmoplastic extracellular matrix in order to permit cell studies under in vivo-like conditions. This Unit provides a protocol and troubleshooting guide for the preparation of high density fibrillar collagen (HDFC) matrices that closely model the desmoplastic collagenous matrix of malignant tumors. It then describes the use of this HDFC matrix for in vitro cell studies of invadopodia formation and function in extracellular matrix invasion. In addition, this Unit also provides a detailed protocol for immuno-labeling of invadopodial proteins and detection of HDFC matrix degradation associated with invadopodia to permit invadopodia visualization using fluorescence microscopy.