Can you recommend an alternative to the typical 4-day assay workflow for using iCell Cardiomyocytes2 in MEA and xCELLigence applications?

We developed an alternative workflow using iCell® Cardiomyocytes2 with 48-well MEA (Axion Maestro) or 96-well xCELLigence RTCA Cardio (ACEA Biosciences) platforms that provides additional assay flexibility outside a typical 5-day work week. Using this alternative schedule, you can still thaw cells during the week and perform assays within the recommended day 4 through 8 post-thaw timeframe without having to change the medium over the weekend. Note that it is very important to adhere to the recommended seeding days and culture conditions, as we have not deviated outside of this workflow range.

Alternative Timeline (solely applicable for Axion MEA (48-well plate) and ACEA xCELLigence RTCA Cardio systems:

  • Wednesday (day 0): Thaw and plate cells according to MEA or xCELLigence Application Protocol.
  • Wednesday – Friday (days 0 – 2): Culture and maintain cells for first 2 days post-plating according to MEA or xCELLigence Application Protocol. This includes a 100% medium change (iCell Cardiomyocytes Maintenance Medium) toward the end of the workday on Friday.
  • Saturday – Sunday (days 3 – 4): Incubate cells at 37°C in 5% CO2 over the weekend without a medium change.
  • Monday (day 5): Perform a 100% medium change early on Monday morning.
  • Tuesday – Wednesday (days 6 – 7): Perform a 100% medium change either the day before the assay (Tuesday evening) or on the day of the assay (Wednesday morning, allowing at least 4 hours for cells to recover from the medium change before performing the assay).

Cell performance of the cells in the standard 4-day (M - F) and 7-day (W – W) protocol were very similar, as measured by our standard baseline parameters on MEA (beat rate, amplitude, FPD) and xCELLigence (beat rate, amplitude, beating rhythm irregularity) as described in the Application Protocols. Data has also been generated by ACEA on the xCELLigence CardioECR (see link to CardioECR solution) showing that the 7-day workflow also works well on that platform (data not shown).

For more information about our cardiomyocyte protocols for electronic microplates like CardioECR (ACEA Bioscience), see Solution 00000290A.


  1. Application Protocol: Measuring Cardiac Electrical Activity: Field Potential Detection the Maestro Multielectrode Array
  2. Application Protocol: Measuring Cardiac Activity: Impedance Detection with xCELLigence RTCA Cardio System