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. 2009 Jul 20:(29):1475.
doi: 10.3791/1475.

An in vitro FluoroBlok tumor invasion assay

Jeff Partridge  1 , Paula Flaherty
Affiliations

An in vitro FluoroBlok tumor invasion assay

Jeff Partridge et al. J Vis Exp. .

Abstract

The hallmark of metastatic cells is their ability to invade through the basement membrane and migrate to other parts of the body. Cells must be able to both secrete proteases that break down the basement membrane as well as migrate in order to be invasive. BD BioCoat Tumor Invasion System provides cells with conditions that allow assessment of their invasive property in vitro(1,2). It consists of a BD Falcon FluoroBlok 24-Multiwell Insert Plate with an 8.0 micron pore size PET membrane that has been uniformly coated with BD Matrigel Matrix. This uniform layer of BD Matrigel Matrix serves as a reconstituted basement membrane in vitro providing a true barrier to non-invasive cells while presenting an appropriate protein structure to study invasion. The coating process occludes the pores of the membrane, blocking non-invasive cells from migrating through the membrane. In contrast, invasive cells are able to detach themselves from and migrate through the coated membrane. Quantitation of cell invasion can be achieved by either pre- or post-cell invasion labeling with a fluorescent dye such as DiIC(12;)(3) or calcein AM, respectively, and measuring the fluorescence of invading cells. Since the BD FluoroBlok membrane effectively blocks the passage of light from 490-700 nm at >99% efficiency, fluorescently-labeled cells that have not invaded are not detected by a bottom-reading fluorescence plate reader. However, cells that have invaded to the underside of the membrane are no longer shielded from the light source and are detected with the respective plate reader. This video demonstrates an endpoint cell invasion assay, using calcein AM to detect invaded cells.

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References

    1. Sethi G, Ahn KS, Pandey MK, Aggarwal BB. Celastrol, a novel triterpene, potentiates TNF-induced apoptosis and suppresses invasion of tumor cells by inhibiting NF-κB-regulated gene products and TAK1-mediated NF-ΚB activation. Blood. 2007;109:2727–2735. - PubMed
    1. Takada Y, Kobayashi Y, Aggarwal BB. Evodiamine Aboloishes Constitutive and Inducible NF-κB Activation by Inhibiting IκBα Kinase Activation, Thereby Suppressing NF-κB-regulated Antiapoptotic and Metastatic Gene Expression, Up-regulating Apoptosis, and Inhibiting Invasion. J. Biol. Chem. 2005;280:17203–17212. - PubMed
    1. Harikumar KB, Kunnumakkara AB, Ahn KS, Anand P, Krishnan S, Guha S, Aggarwal BB. Modification of the cysteini residues in IκBα kinase and NF-κB (p65) by xanthohumol leads to suppression of NF-κB-regulated gene products and potentiation of apoptosis in leukemia cells. Blood. 2009:113–2003. - PMC - PubMed
    1. Nair AS, Sishodia S, Ahn KS, Kunnumakkara AB, Sethi G, Aggarwal BBDeguelin. an Akt Inhibitor, Suppresses IΚBα Kinase Activation Leading to Suppression of NF-κB-Regulated Gene Expression, Potentiation of Apoptosis, and Inhibition of Cellular Invasion. J. Immunol. 2006:177–5612. - PubMed
    1. Partridge J, Qian S. Quantitative and High-Throughput Screening of Tumor Cell Invasion using a Cell-based Model; Society for Biomolecular Screening 2005 Conference; 2005 Sept 11-15; Geneva, Switzerland. 2005.

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