Preparation of High-Density Fibrillar Collagen Matrices That Mimic Desmoplastic Tumor Stroma.

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. © 2016 by John Wiley & Sons, Inc.