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首页> 外文期刊>Current Protocols in Cell Biology >UNIT 10.19 Preparation of High-Density Fibrillar Collagen Matrices That Mimic Desmoplastic Tumor Stroma
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UNIT 10.19 Preparation of High-Density Fibrillar Collagen Matrices That Mimic Desmoplastic Tumor Stroma

机译：UNIT 10.19模仿增生性肿瘤基质的高密度原纤维胶原蛋白基质的制备

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摘要

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 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 matrix for in vitro cell studies of invadopodia formation and function in extracellular matrix invasion. In addition, it provides a detailed protocol for immunolabeling of invadopodial proteins and detection of HDFC matrix degradation associated with invadopodia to permit visualization of invadopodia using fluorescence microscopy.
机译：侵袭性肿瘤的基质由于去增生反应而变得富含致密的原纤维胶原蛋白。这种增生性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来模拟这种增生的细胞外基质，以便在类似体内的条件下进行细胞研究。本单元提供了协议和故障排除指南，用于准备高密度原纤维胶原蛋白（HDFC）基质，该基质可以密切模拟恶性肿瘤的促增生性胶原基质。然后描述了该基质在体外细胞研究侵袭足形成和细胞外基质侵袭功能中的用途。此外，它提供了一个详细的方案，用于对卵小体蛋白进行免疫标记和检测与卵小体相关的HDFC基质降解，以允许使用荧光显微镜对卵小体进行可视化。

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