Which protocol should I use for the ACEA Biosciences xCELLigence platform and your cardiomyocytes?

Our Application Protocol describes how to detect iCell® Cardiomyocytes beat rhythmicity using the xCELLigence RTCA Cardio System (ACEA Biosciences), which is a non-invasive label-free platform that measures impedance changes across the cardiac monolayer to indirectly assess cardiomyocyte viability, contractility, and electrical activity. To reduce workflow steps, our protocol allows thawing and plating of the cells directly onto the E-Plate on the same day. Using this system, you can culture and maintain iCell Cardiomyocytes on an E-Plate for extended durations, enabling measurement of acute and sub-acute drug-induced effects. Analysis options include the ability to compare changes in beat rate, amplitude, and beat rhythm irregularity using ACEA software, and normalizing post-dose recordings to pre-dose recordings taken on the same day to obtain values for beat rate and amplitude.

General protocol recommendations:

  • Assay timing: 14 days post thaw using direct plating into E-Plate
  • Assay density: Plate 20,000 plated cells in 50 μl per well (+ 50 μl medium already in the well)
  • Assay media: iCell Cardiomyocytes Plating Medium and iCell Cardiomyocytes Maintenance Medium
  • Assay ECM: fibronectin (10 μg/ml)

References:

  1. Application Protocol: Measuring Cardiac Activity: Impedance Detection with xCELLigence RTCA Cardio System
  2. Guo et al. (2013) Refining the Human iPSC-Cardiomyocyte Arrhythmic Risk Assessment Model
  3. The xCELLigence System: RTCA SP & MP Instrument Training DVD, ACEA Video