Abstract

2D and 3D High Content Assays of Tumor Cell Invasion

Maria Montoya, Head of Cellomics Unit, National Center for Cardiovascular Research

The tumor microenvironment is defined by stromal cells and ECM components which have important implications in the proliferation, differentiation and invasive capabilities of tumor cells. We have established a 2D cellular assay to study the organization of the extracellular matrix and overall stroma-mediated remodeling of the matrix by high content analysis to explore molecular mechanisms underlying fibroblast-driven cancer progression. Cellular growth in 2D versus 3D differ in mechanical, biochemical, and contextual factors which can affect intracellular signal transduction pathways with important implications in the polarization and differentiation status of cells. Three dimensional cell invasion assay of cancer associated fibroblasts co-culture with tumor cells aimed to identify compounds or molecular targets for inhibiting tumor cell invasion will be presented. HCS assay is based on a novel method that compute tridimensional cell invasion on a cell by cell basis from a cloud of tumor cells obtained from DAPI staining in confocal Z-stacks of 3D images using geometrical information. Approaches to high content 2D and 3D geometries and multiparametric-analysis as well as the address of inherent problems found in the experimental setup will be discussed.