Are your cardiomyocytes compatible with FLIPR and FDSS platforms?
Yes. Our Application Protocol describes how to handle iCell® Cardiomyocytes for use on the FLIPR Tetra High-Throughput Screening System (Molecular Devices) and provides basic instructions for compound treatment, data acquisition and analysis. We demonstrated culture of iCell Cardiomyocytes on 96- or 384-well plates, where they formed a stable, electrically and mechanically active syncytium amenable to measuring drug-induced perturbations for in vitroscreening of compound efficacy and toxicity. Others have demonstrated the utility of the FDSS/μCELL system (Hamamatsu Photonics) with our iCell Cardiomyocytes in posters 5,6 and in the peer-reviewed publication by Puppala et al. Additionally, we have also developed an Application Protocol for using iCell Cardiomyocytes2 on the FDSS/μCELLsystem.
For both of these screening systems, assay timing is critical, and we recommend that you adhere to the workflow in the application protocol. Ideal timing for FLIPR analysis of iCell Cardiomyocytes is on day 14 post-thaw, which shows consistent and uniform results across wells when measuring beat rate and dose response. To reduce the potential for calcium dye toxicity, we recommend that you screen on the same day that you add calcium dye.
It is also important to pay attention to temperature with cardiomyocytes, as deviations from a controlled environment can reduce the beat rate of the cells. To overcome temperature variations, always return plates to 37°C incubator as soon as possible during incubation steps.
General protocol recommendations:
- Assay timing: 14 days post plating
- Assay density (direct plating): Plate 5,000 cells in 40 μl per well in a 384-well plate or 20,000 cells in 100 μl in a 96-well plate
- Assay media: iCell Cardiomyocytes Maintenance Medium
- Assay ECM: 0.1% Gelatin
For more information about optimizing culture conditions for iCell Cardiomyocytes, including edge effects and environmental issues, see solution suggestions here: 00000043E
- Application Protocol: Measuring Cardiac Activity: Intracellular Calcium Flux Detection on the FLIPR Tetra System
- Application Note (Molecular Devices): EarlyTox Cardiotoxicity Kit Provides Biologically Relevant Cardiotoxicty Data Earlier in the Drug Discovery Process
- Sirenko et al, 2013 Multiparameter In Vitro Assessment of Compound Effects on Cardiomyocyte Physiology Using iPSC Cells
- Application Procotol: Measuring Cardiac Activity: Intracellular Calcium Flux Detection with FDSS/μCELL
- Poster Presentation: Saito et al. (SLAS 2014) Electric Field Stimulation of iPSC-Derived Cardiomyocytes Using Hamamatsu FDSS/μCELL with Fast Data Acquisition
- Poster Presentation: Roman et al. (SPS 2014) Determination of Pro-arrhythmic Effects of Compounds in Human iPSC-Derived Cardiomyocytes Using FDSS/μCell Imaging Platform
- Puppala et al. 2013 Comparative Gene Expression Profiling in Human-Induced Pluripotent Stem Cell–Derived Cardiocytes and Human and Cynomolgus Heart Tissue
- Bedut et al. 2016 High-throughput Drug Profiling with Voltage- and Calcium-Sensitive Fluorescent Probes in Human iPSC-Derived Cardiomyocytes