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P2056
Division Arrested Cells Simplifying Assay Design & Increasing Flexibility
Presenter Steven Murphy, Invitrogen Discovery Sciences, USA
Additional Authors: Sherry Boozer, Michael Bagguley, Amanda Hibbard, Corey Cooper, Bonnie Hanson, and Mei Cong
Cellular assays are valuable tools for both the identification and characterization of drug candidates but their use has been limited by the long lead times required to expand cells and the high labor costs of maintaining cells in culture. Division arrested cells provide a means to over come these barriers and allow cellular assays to resemble biochemical assays in terms of flexibility and cost without sacrificing content. The advantages of division arrested cells are especially apparent in counter screening environments where screening is done on a periodic basis against a large panel of cellular assays. Here we show that division arrest technology is broadly applicable to a wide range of cell lines including those expressing GPCRs, nuclear receptor and ion channel cell lines. The correlation between EC50 values and Z’ values determined in division arrested cells and dividing cells will be shown for 81 GPCR cells lines and 20 nuclear receptor cell lines. The process of division arresting cells can be used to simplify complex assays so that they are more appropriate for an HTS environment. For example, nuclear receptor cell-based assays that require in the plate serum starvation followed by an aspiration step can be simplified by using division arrested cells that are serum starved prior to the division arrest process. The division arrested nuclear receptor cells can be directly used in the assay thereby eliminating the in plate serum starvation and aspiration. In addition, we have found that division arrested cells can go through multiple freeze/thaw cycles without losing activity, which increases flexibility in experimental design and reduces reagent costs. Division arrested cells are functionally comparable to dividing cells, decrease labor costs, increase flexibility, and allow assays to be simplified such that complex assays are more amendable to high-throughput screening.