Notes
iCell Cardiomyocytes User’s Guide
11
lot/shipment. Using the lot number, you can also look up your Certificate of
Analysis online at www.cellulardynamics.com/coa/.
Plating iCell Cardiomyocytes in 96-well Cell Culture Plates
for Cell-based Assays
The recommended plated density for performing cell-based assays in 96-well cell
culture plates using iCell Cardiomyocytes is 20,000 plated cells/well. However, as
mentioned in the previous section, the optimal plated density is dependent on the
biology in question as well as the sensitivity of the assay and must be determined
empirically.
The following procedure describes how to plate 20,000 cells/well in a 96-well cell
culture plate assuming a seeding volume of 100
μ
l/well, a
Plating Efficiency
of
45%, a post-thaw
Viable Cell Density
of 0.49 x 10
6
cells/ml, and an actual post-
thaw cell volume of 9.5 ml (0.5 ml was used to attain the
Viable Cell Density
).
This same procedure can be used to plate cells in other culture vessel formats by
substituting the appropriate recommended cell numbers and seeding volumes.
1.
Prepare the thawed cell suspension for plating into a 96-well cell culture plate.
a. Obtain
the
Plating Efficiency
from the Certificate of Analysis. Alternatively,
use the lot number to look up your Certificate of Analysis online at
www.cellulardynamics.com/coa/ (for purposes of this example, the
Plating
Efficiency
is 45%).
b. Obtain
the
Viable Cell Density
(cells/ml) from the Determining Viable Cell
Density section earlier in this chapter (for purposes of this example, the
Viable Cell Density
is 0.49 x 10
6
cells/ml).
c. Calculate
the
Target Plating Density
:
/
/
For purposes of this example, the desired number of plated cells/well is
20,000, and the suggested seeding volume is 100
μ
l. Thus, the
Target
Plating Density
is
0.2 10
/
20,000
/
0.1
d. Calculate
the
Total Plating Volume
necessary to bring the thawed cell
suspension to the
Target Plating Density
.
⁄
⁄
where
Actual Volume
is the post-thaw suspension volume after cell
counting. Thus, the
Total Plating Volume
is
10.47
9.5
0.49 10
⁄
0.45
0.2 10
⁄
e. Add sufficient Plating Medium to the thawed cell suspension to achieve the
calculated
Total Plating Volume
.
