Promega Cell Viability Assay Designed Specifically for 3D Microtissue Culture
11 Jun 2014
Promega Corporation launches a new assay validated for assessing the viability of cells in three-dimensional (3D) microtissue model systems. CellTiter-Glo® 3D Cell Viability Assay penetrates large 3D microtissue samples to provide more accurate determination of viable cells. The assay has been validated on a range of 3D microtissue model systems, generated using scaffold and scaffold-free systems. CellTiter-Glo 3D Assay uses the robust Ultra-Glo™ Recombinant Luciferase in a solution optimized to penetrate denser 3D microtissues for a complete release of ATP and accurate assessment of viability by luminescence detection.
Scientists have found that 3D microtissue cell culture models better reproduce the cell-to-cell and cell-to-matrix interactions occurring in living organisms, providing a more predictive in vitro experimental model system. Most, if not all commercial cell-based assays, including MTT, resazurin and current ATP assays, were developed using 2D monolayer models, are less effective on denser 3D microtissue models and cannot fully penetrate larger 3D microtissue spheroids.
"There is a clear need for cell viability assays validated for use in 3D microtissue culture. The novel chemistry of CellTiter-Glo 3D Reagent and the optimized protocol are designed specifically for these 3D model systems," says Dr. Terry Riss, Senior Product Specialist at Promega.
The CellTiter-Glo 3D Assay has a simple add-mix-incubate-measure protocol that is completed in 30 minutes or less. The reagent includes optimized detergent concentration, ATPase inhibitors for protection of released ATP, proprietary thermostable Ultra-Glo™ Recombinant Luciferase and luciferin. CellTiter-Glo 3D Reagent acts on 3D microtissue cultures to release ATP, which is then consumed in the luciferase reaction to produce a stable glow-type luminescence. The amount of light produced correlates with the number of viable cells in culture. The simple protocol and high sensitivity of the assay make it amenable to higher throughput screening applications.
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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.