Abstract 5006: A 3-dimensional cell invasion assay compatible with high content screening

Abstract

Abstract The transition from non-invasive phenotype to invasive phenotype of tumor cells marks the switch from a benign tumor that can be successfully treated via surgery to a more malignant form of cancer. Understanding the mechanisms underlying this hallmark event, which enables tumor cells to invade through extracellular matrix, is critical for discovering pathways and new targets to develop anti-metastatic strategies. Previously, we developed a cell based assay for the study of cell migration in 96-well plates. A self-dissolving biocompatible gel (BCG) is utilized to form uniformly sized, cell-free detection zones in collagen I coated 96-well plates. When cells are seeded into these wells, they pattern in an annular monolayer surrounding the BCG. Once the BCG dissolves, an overlay of collagen I is applied to the assay wells, following which cells can invade in 3-dimensions into the detection zone previously occupied by the BCG. This “Oris™ Pro” cell invasion assay presents a straightforward, accessible and quantifiable cell method to study cell invasion, and can be used for example to evaluate candidate drugs targeting tumor invasion. Microscopic visual assessment of cell invasion in the presence of inhibitors is made by use of cell stains at multiple time-points. In this study we demonstrate the use of this assay platform for robust and reproducible analysis of HT-1080 and MDA- MB-231 cell invasion in 3-D. The presentation will discuss (i) factors that influence cell invasion, including density of collagen overlay and time; (ii) microscopic monitoring of cell invasion in the presence of potential inhibitors throughout the duration of the experiment; and (iii) the qualitative and quantitative readouts that can be obtained using this 3-dimensional cell invasion assay. Citation Format: Jennifer A. Fronczak, Matthew S. Gajeski, Erica L. Beckman, Laura Vollmer, Andreas Vogt, Gopal Krishnan. A 3-dimensional cell invasion assay compatible with high content screening. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5006. doi:10.1158/1538-7445.AM2014-5006