Abstract 4137: An improved assay system for cell migration and invasion
| Autor: | Paula Flaherty, Stephen Rimsa, Jeff Partridge |
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| Rok vydání: | 2013 |
| Předmět: | |
| Zdroj: | Cancer Research. 73:4137-4137 |
| ISSN: | 1538-7445 0008-5472 |
| Popis: | In vitro cell migration and invasion assays are frequently used as model systems for quantifying the directed movement of cells towards a chemoattractant stimulus, or to measure how a particular drug, antibody or extra cellular matrix (ECM) coating affects that movement. Analysis of this movement is often accomplished through the use of visual methods that are either time-consuming, labor-intensive, or subjective. To address these issues, an improved fluorescence blocking version of the Boyden chamber was developed. As cells migrate or invade through a unique fluorescence blocking microporous membrane, they can be detected using a bottom-reading spectrofluorometer or fluorescence microscope. Cells remaining in the upper chamber of the insert are shielded from this detection, allowing for quantitation of cell numbers in a homogeneous assay system. The technology blocks ≥ 98% transmission of light from 360 - 690 nm as measured by transmission spectrophotometry, and improvements have been made over previous embodiments such that >50% more light in the range of 360 - 490 nm is blocked. These enhancements now permit the use of the blue fluorescent Hoechst dyes and DAPI for direct cell enumeration and multiplexing. Functional applications of this technology were demonstrated in several well-established assays: endpoint staining in a model tumor invasion system yielded a Z’-factor >0.5; real-time chemotaxis of monocytes demonstrated a peak signal between 25-35 minutes; and migration of human endothelial cells to VEGF was demonstrated with high reproducibility (CV < 15%). Additionally, the measured IC50 values of chemotherapeutic compounds including paclitaxel and doxorubicin remained consistent with previously reported results. FluoroBlok™ cell culture inserts can efficiently be used for high-throughput cell migration studies, are automation friendly and yield highly reproducible results. Citation Format: Jeff Partridge, Stephen Rimsa, Paula Flaherty. An improved assay system for cell migration and invasion. [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 4137. doi:10.1158/1538-7445.AM2013-4137 |
| Databáze: | OpenAIRE |
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