The CellTiter-Glo® 3D Assay is a simple assay: Add a single reagent to cells in medium and quantitate cell viability of 3D microtissues in 30 minutes. This luminescent assay is scalable for use in high-throughput screening, is compatible with many 3D culture methods and offers greater sensitivity than colorimetric and fluorescentbased cell viability assays. A. HCT116 Human Cancer Cell Microtissues 1,000 10,000 1,000 100 10 10 1 CellTiter-Glo® 3D alamarBlue® MTT 1 CellTiter-Glo® 3D alamarBlue® MTT Figure 1. CellTiter-Glo® 3D Assay provides better signal-to-background ratios than colorimetric or fluorometric assays. Panel A. HCT116 colon cancer cells seeded into an InSphero GravityPLUS™ 96-well hanging-drop platform and grown to generate ~340μm spheroids. Panel B. InSphero Insight™ human liver microtissues (~250μm). All microtissues were assayed using the CellTiter-Glo® 3D, alamarBlue® and MTT assays according to manufacturer’s protocols. Total assay times for the CellTiter-Glo® 3D, alamarBlue® and MTT assays were 30 minutes, 3 hours and 8 hours, respectively. A. Hanging Drop Method 120 100 80 60 40 20 0 –10 –9 –8 –7 Log [Compound], M C. Matrigel® 120 100 80 60 40 20 0 Figure 2. CellTiter-Glo® 3D Assay is compatible with a variety of 3D culture methods. HCT116 colon cancer cells were seeded as follows: 400 cells in hanging-drop (Panel A); 1,000 cells in ultra-low attachment (ULA) or Matrigel® (Panels B and C). Microtissues were grown for 4 days, treated with compounds for 48 hours, then assayed with the CellTiter-Glo® 3D Reagent. Luminescence was recorded at 30 minutes. –10 –9 –8 –7 Log [Compound], M enews | April 2015 –6 –5 –4 –6 –5 panobinostat terfenidine trichostatin A B. ULA –4 120 100 80 60 40 20 0 100 B. Human Liver Microtissues –10 –9 –8 –7 Log [Compound], M –6 –5 –4 Percent Relative Activity Percent Relative Activity Signal-to-Background Ratio Signal-to-Background Ratio 12386MB Percent Relative Activity 12331MC Pagina 4
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Promega Benelux - Monthly online Magazine - April 2015 edition Lees publicatie 100Home