Cardiac Spheroids on SpheroHD Protocol

Preparing the SpheroHD Chip

1. Pipette 10 μl of fibronectin solution (50 µg/mL in PBS) into each of the four chambers of the Ibidi insert on the SpheroHD chip.
2. Incubate the FN-coated SpheroHD chip in a cell culture incubator at 37°C, 5% CO₂ for at least 60 minutes.

Culturing and Plating Cardiac Spheroids

1. Aspirate FN from the chambers and add 10 µl of cardiac spheroid media to each
   chamber of the Ibidi insert on the SpheroHD chip.

   Tip: More information on proper handling of the Ibidi 4 well FulTrac mico insert can be found on Ibidi’s website (Cat. No: 80486).

2. Centrifuge at 1500 x g for 1 minutes using the provided single-well centrifuge holder to remove bubbles. Remove the media from each chamber.

3. Using a 200 µl wide-bore pipette tip, add one cardiac spheroid from culture to each chamber of the Ibidi insert on the SpheroHD.

   Tip: Transferring spheroids in small volumes (10 µl, e.g.) can be difficult. Spheroids can be transferred in larger volumes by pipetting up the spheroid, allowing it to fall to the bottom of the wide bore tip, and touching the tip to media in the chamber to transfer the spheroid.

4. Centrifuge at 100 x g for 1 minute using the provided single-well centrifuge holder to bring the spheroids to the bottom of the chambers. Check under the microscope to ensure that all spheroids are positioned over the electrodes at the bottom of the well. Repeat or increase speed if necessary.
5. Once the spheroids are in place, incubate the SpheroHD MEA chip in a cell culture incubator at 37°C, 5% CO₂ for 1 hour.
6. Add enough cardiac spheroid media (about 1.75 mL) to cover the top surface of the Ibidi insert. Maximum volume for the SpheroHD chip is 2 mL.
7. Take recordings using the MEA Creator Kit on the Maestro Pro, Edge, or Volt as desired.

Required Materials

Consumables

Equipment