球体
细胞外基质
肿瘤微环境
蛋白酵素
癌细胞
转移
癌相关成纤维细胞
分泌物
细胞
体内
细胞生物学
化学
癌症研究
基质(化学分析)
活体细胞成像
癌症
生物
肿瘤细胞
体外
生物化学
酶
生物技术
遗传学
色谱法
作者
M. Ángeles Villaronga,Saúl Álvarez–Teijeiro,Francisco Hermida‐Prado,Marta Garzón-Arango,Victoria Sanz‐Moreno,Juana M. García–Pedrero
出处
期刊:Methods in molecular biology
日期:2018-01-01
卷期号:: 145-154
被引量:5
标识
DOI:10.1007/978-1-4939-7595-2_14
摘要
Tumor growth and progression is the result of a complex process controlled not only by malignant cancer cells but also by the surrounding tumor microenvironment (TME). Cancer associated fibroblasts (CAFs), the most abundant cellular component of TME, play an active role in tumor invasion and metastasis by promoting cancer cell invasion through cell-cell interactions and secretion of pro-invasive factors such as extracellular matrix (ECM)-degrading proteases. Due to their tumor-promoting activities, there is an emerging interest in investigating CAFs biology and its potential as drug targets for cancer therapies. Here we describe an easy and highly reproducible quantitative method to analyze CAF invasive activity by forming multicellular spheroids embedded into a three-dimensional (3D) matrix that mimics in vivo ECM. Subsequently, invasion is monitored over time using a time-lapse microscope. We also provide an automated image analysis system that enables the rapid quantification of the spheroid area increase (invasive area) over time. The use of a 96-well plate format with one CAF spheroid per well and the automated analysis provides a method suitable for drug screening test, such as protease inhibitors.
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