A new technology for in vitro chemotaxis assays (CAM4P.159)

• Published in: The Journal of immunology (1950) 2015-05, Vol.194 (1_Supplement), p.185-185.17
• Main Authors: APPLEDORN, DANIEL, O'Clair, Lindy, Tilhonenko, Maria, Roddy, Meagan, Schroeder, Kirk
• Format: Article
• Language: English
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.194.Supp.185.17

Chemotactic-driven cell migration is an important biological process in many areas of life science research including immunology, cancer and developmental biology. The dominant high-throughput (96-well format) solution for measuring cellular chemotaxis in vitro is the Boyden-chamber. However, Boyden chambers have some inherent liabilities including: a) the inability to directly visualize the cell migration process, b) a requirement of tens of thousands of cells per well, c) the need to label cells for quantitation, d) a relative insensitivity to surface integrin signaling, and e) high random migration artifacts. Microfluidic solutions have sought to eliminate some of these deficiencies, specifically by providing the ability to visualize the cell migration process, but at the expense of relatively low throughput and cumbersome quantitation methods. We have developed a new technology which eliminates most of the deficiencies of existing Boyden chambers and most importantly, provides a direct visualization of the migration process as it occurs. Integrated metrics precisely quantify the chemotactic response using significantly fewer cells (500-5000 cells per well) in a 96-well format. This assay method does not require labeling for quantitation, has a kinetic read-out, and is sensitive to surface-integrin signaling. Pharmacological data examples from T cells, neutrophils, cancer cells, and endothelial cells will be shown with observations enabled by the real-time visualization.