What general assay guidelines do you have for cardiomyocytes?
You can perform most assays after iCell® Cardiomyocytes re-animate from cryopreservation. This re-animation usually occurs about 7 days post-plating, after a confluent monolayer forms and establishes synchronous beating. Cell-beating assays (e.g. impedance and MEA) may benefit from a longer culture of 10 - 14 days. Note that cells are post-mitotic and will not proliferate in culture.
Plating Density
- Plate cells to enable formation of a confluent monolayer whenever possible.
- Density can influence gene expression.
Extracellular Matrix (ECM)
- iCell Cardiomyocytes work with a variety of ECMs.
- See Solution 00000082A, below, for details about using gelatin and other ECMs.
Media
- iCell Cardiomyocytes work with a variety of media.
- Note that ionic concentrations are important for cell beating.
- Avoid calcium-free media.
Timeline
- Do not perform assays on freshly thawed (within 5 days) or dissociated cardiomyocytes (within 2 days) whenever possible.
- For beat rhythm assays, we recommend using cells cultured for 10 - 14 days.
For biochemical and toxicity assays, we recommend using cells cultured for 7 days.
Reference:
