Ray Arja, Morford Rachel K, Provenzano Paolo P
Department of Biomedical Engineering, University of Minnesota, Minneapolis, Minnesota.
University of Minnesota Physical Sciences in Oncology Center, Minneapolis, Minnesota.
Curr Protoc Stem Cell Biol. 2018 Aug;46(1):e57. doi: 10.1002/cpsc.57. Epub 2018 May 24.
Cell migration is strongly influenced by the organization of the surrounding 3-D extracellular matrix. In particular, within fibrous solid tumors, carcinoma cell invasion may be directed by patterns of aligned collagen in the extra-epithelial space. Thus, studying the interactions of heterogeneous populations of cancer cells that include the stem/progenitor-like cancer stem cell subpopulation and aligned collagen networks is critical to our understanding of carcinoma dissemination. Here, we describe a robust method to generate aligned collagen matrices in vitro that mimic in vivo fiber organization. Subsequently, a protocol is presented for seeding aligned matrices with distinct carcinoma cell subpopulations and performing live cell imaging and quantitative analysis of cell migration. Together, the engineered constructs and the imaging techniques laid out here provide a platform to study cancer stem cell migration in 3-D anisotropic collagen with real-time visualization of cellular interactions with the fibrous matrix. © 2018 by John Wiley & Sons, Inc.
细胞迁移受到周围三维细胞外基质组织的强烈影响。特别是在纤维状实体瘤中，癌细胞的侵袭可能由上皮外空间中排列的胶原蛋白模式所引导。因此，研究包括干细胞/祖细胞样癌症干细胞亚群和排列的胶原网络在内的异质性癌细胞群体之间的相互作用，对于我们理解癌症扩散至关重要。在这里，我们描述了一种在体外生成模拟体内纤维组织的排列胶原基质的强大方法。随后，给出了一个方案，用于将不同的癌细胞亚群接种到排列的基质上，并对细胞迁移进行活细胞成像和定量分析。总之，本文阐述的工程构建体和成像技术提供了一个平台，用于研究癌症干细胞在三维各向异性胶原中的迁移，并实时可视化细胞与纤维基质的相互作用。© 2018约翰威立父子出版公司
