Compound Profiling for Anti-Cancer Activity using NCI-60 Cell Lines and Tecan’s Fluent Laboratory Automation Solution

19 Jul 2015

Tecan has re-invented automation with Fluent, a unique instrumentation concept built around the application-specific needs of cell biology and drug discovery laboratories. Fluent breaks new ground, delivering greater capacity and increased speed; the platform provides superior throughput and walk-away time, making it easier to get more done, more effectively. This application notes describes the successful automation of a cell viability assay using a selection of NCI cell lines which are commonly employed in cancer drug discovery.

CellTiter-Glo® 3D Cell Viability Assay

Promega Corp.

The CellTiter-Glo® 3D Cell Viability Assay is a homogeneous method to determine the number of viable cells in 3D cell culture based on quantitation of the ATP present, which is a marker for the presence of metabolically active cells.   This ready-to-use reagent is based on the original CellTiter-Glo® Luminescent Cell Viability Assay chemistry and eliminates the need to combine buffer with lyophilized substrate when preparing reagent. The CellTiter-Glo® 3D Cell Viability Assay is formulated with more robust lytic capacity and is designed for use with microtissues produced in 3D cell culture. The homogeneous assay procedure involves addition of a single reagent (CellTiter-Glo® 3D Reagent) directly to cells cultured in serum-supplemented medium. Cell washing, removal of medium and multiple pipetting steps are not required. This assay is compatible with multiwell-plate formats, making it ideal for automated high-throughput screening (HTS). The CellTiter-Glo® 3D Assay has been used successfully with 3D microtissue cell culture produced via hanging-drop plates, ultra-low attachment plates, Matrigel®-coated plates, agarose-coated plates, cultures suspended in methylcellulose and Alvetex® plates. Features & Benefits:   Robust Penetration into Microtissues - Improved lytic capacity allows use over a broad range of microtissue sizes compared to other viability assay methods. Ready-to-Use Reagent - No mixing of components required; simply thaw, equilibrate to room temperature and "add-mix-incubate-measure". Convenient for HTS applications. Fast Results - Data can be recorded in 30 minutes or less after adding reagent, quicker than when using colorimetric or fluorometric viability assays. Superior Sensitivity - The signal-to-background ratio of this assay applied to microtissues is much greater than that of standard colorimetric and fluorometric assays. Flexible Format - The assay can be used with various multiwell formats (96-well and regular or low-volume 384-well). Data can be recorded by luminometer, CCD camera or other imaging devices capable of reading luminescence in multiwell plates. Glow-Type Signal - Stable luminescent signal half-life >3 hours, depending on cell type and culture medium used, allows batch mode or consecutive processing of multiple plates. Applications: Cell viability assays with 3D microtissue cell culture or standard cell culture samples. Cytotoxicity assays with 3D microtissue cell culture or standard cell culture samples.

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Compound Profiling for Anti-Cancer Activity using NCI-60 Cell Lines and Tecan’s Fluent Laboratory Automation Solution