8
Cellular Dynamics International, Inc.
Notes
5.
Rinse the empty iCell Cardiomyocytes cryovial with 1 ml of room temperature
Plating Medium to recover any residual cells from the vial. Transfer the 1 ml
Plating Medium rinse from the cryovial drop-wise over 90 seconds (i.e. 1 drop
every 4 - 5 seconds) to the 50 ml centrifuge tube containing the iCell
Cardiomyocytes cell suspension. Gently swirl the tube while adding the
medium to mix the solution completely and minimize the osmotic shock on the
thawed cells.
6.
Slowly add 8 ml (3.5 ml for 0.5 unit size of cardiomyocytes) of room
temperature Plating Medium to the 50 ml centrifuge tube. Add the first 1 ml
drop-wise over 30 - 60 seconds. Then add the remaining volume over the next
~30 seconds. Gently swirl the centrifuge tube while adding the medium.
7.
Gently mix the contents of the 50 ml centrifuge tube by inverting 2 - 3 times.
Gentle mixing is critical to ensure maximum viability. Avoid vigorous shaking or
vortexing of the cell suspension.
Note
: Thaw up to 3 vials of iCell Cardiomyocytes at one time. Once thawed,
you can pool the contents of the vials before adding the rinse and final volume
of Plating Medium. Follow the timing outlined in steps
5
and 6. For example, if
pooling 3 vials, add each 1 ml of rinse over 90 seconds (270 seconds total).
Drop-wise addition of Plating Medium to the cell suspension is critical
to minimize osmotic shock and ensure maximum viability and
attachment of the cells to the plating substrate. See the Handling
iCell Cardiomyocytes Training Video available online at
www.cellulardynamics.com/cm handling/.
It is critical to add the Plating Medium slowly to ensure maximum viability
and attachment of the cells once plated. See the Handling iCell
Cardiomyocytes Training Video available online at
www.cellulardynamics.com/cm_handling/.
