Abstract 4897: A robust 384-well cell migration assay for high content analysis (HCA) of cells treated with anti-cancer therapeutics

Abstract

Abstract Platypus Technologies has developed a first-in-class 384-well cell migration assay to enable high throughput screening (HTS) of anti-cancer compounds and wound healing agents on adherent tumor and endothelial cell lines. The assay is completely automatable with liquid handling systems and high content analysis (HCA) instrumentation. This assay format utilizes a centrally located self-dissolving, non-toxic biocompatible gel (BCG) to form a uniformly sized, cell-free detection zone on tissue culture treated or collagen I coated cell culture surfaces. Cells are seeded into 384-well plates and pattern in an annular monolayer surrounding the BCG. Once the BCG dissolves, cells can migrate into the detection zone previously occupied by the BCG. Dose-dependent inhibition of migration was observed using the actin polymerization inhibitors Cytochalasin D and Latrunculin A, the kinase inhibitors Dasatinib and Sorafenib on HT-1080, and human umbilical vein endothelial cells (HUVECs), and the phosphatase inhibitors NSC95397 and BCI on MDA-MB231 human breast cancer cells. While migration of cells was faster on collagen coated surfaces than on tissue culture treated surfaces, the IC 50 values obtained for the compounds and rank order of potency were similar on both surfaces. This assay format allows an unobstructed view of cell motility throughout the duration of the experiment. Cells may be fixed and treated with multiple stains, including Hoechst or DAPI to visualize nuclei and TRITC-phalloidin to observe F-actin, to enable flexible data capture by either enumerating migrating cells or by calculating the area of closure within the detection zone. The 384-well assay was successfully automated using a Biomek 2000 robotic liquid handler and a Cellomics ArrayScan II high content imaging (HCI) instrument. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4897. doi:10.1158/1538-7445.AM2011-4897