Abstract 3799: HTS performance and high-content analysis of antimigratory compound phenotypes using the Oris Pro 384 cell migration assay.

Abstract

Abstract The discovery of anti-metastatic agents that inhibit cancer cell motility has been hindered by a dearth of high-throughput screening (HTS) -compatible cell motility assays. The Oris™ Pro 384 well cell migration assay, developed by Platypus Technologies, is an innovative cell motility assay designed to enable HTS of potential anti-cancer compounds and wound healing agents on adherent tumor and endothelial cell lines. The assay utilizes a centrally located, non-toxic, biocompatible gel (BCG) to form a uniformly sized, cell-free detection zone into which cells migrate. The assay is logistically simple and does not require any mechanical processing steps, such as cell wounding or removal of physical barriers. The assay is fully compatible with laboratory automation, including robotic liquid handlers, plate washers, and high-content screening (HCS) readers. A formal investigation of the accuracy, robustness, and HTS performance of the assay was conducted following guidelines in place at the University of Pittsburgh Drug Discovery Institute (UPDDI). Using MDA-MB-231 human breast cancer cells, the assay was optimized for migration kinetics, cell seeding density, and DMSO tolerance. In multi-day variability studies, the assay delivered signal-to-background ratios > 10 and Z-factors above 0.5 on three consecutive days, was insensitive to process errors or edge-effects, and passed UPDDI Assay Protocol Approval Committee (APAC) criteria. The assay delivered equal HTS performance on two different HCS platforms – the Thermo Scientific Cellomics ArrayScan II and the Molecular Devices ImageXpress®Ultra point scanning confocal reader. Screening of a test cassette of 1280 compounds with known biological activities identified several agents with mechanisms associated with cell motility. The multiparametric nature of the HCS assay permitted correlation of antimigratory activity with cellular toxicity. Our results validate the Oris™ Pro 384 cell migration assay as a logistically simple and information-rich cell motility analysis platform that meets the demands of large scale compound screening. Citation Format: Jennifer Fronczak, Laura Vollmer, Gopal Krishnan, Andreas Vogt. HTS performance and high-content analysis of antimigratory compound phenotypes using the Oris Pro 384 cell migration assay. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3799. doi:10.1158/1538-7445.AM2013-3799