6
Cellular Dynamics International, Inc.
Notes
Chapter 3. Preparing Cell Culture Surfaces
iCell Cardiomyocytes will plate and function on a variety of substrates including
freshly prepared gelatin and fibronectin, which have been shown to support
attachment, viability, and function of iCell Cardiomyocytes with similar efficiencies.
Coating plates with a 0.1% gelatin solution is an economical, simple, and
recommended method for preparing cell culture plates for culturing iCell
Cardiomyocytes.
CDI provides application protocols and notes that recommend assay-specific
substrates. See www.cellulardynamics.com/lit/ for a list of available application
protocols and notes for iCell Cardiomyocytes. Regardless of the substrate of
choice, prepare plating surfaces before thawing iCell Cardiomyocytes.
Preparing 0.1% (w/v) Gelatin Solution
1.
Prepare bottles for mixing the 0.1% gelatin solution.
Do not use bottles that have previously contained or been exposed to
detergent.
Acid-wash new bottles before use.
Use only high-purity, 18 M
sterile water for washing bottles.
2.
Prepare 0.1% gelatin solution by dissolving 500 mg of gelatin (Type A, powder,
cell culture tested) in 500 ml of sterile water in each bottle.
3.
Sterilize the gelatin solution by autoclaving using liquid cycle. The sterile 0.1%
gelatin solution is stable for 3 months when prepared as described above and
stored at room temperature.
Preparing the Gelatin Cell Culture Vessel
1.
Select the cell culture vessel appropriate for your experimental use. Add the
volume of 0.1% gelatin solution specified in the table below. Scale volumes
appropriately for other vessel formats.
Culture
Vessel
Surface Area
(cm
2
)
Volume of 0.1% Gelatin
Solution (ml)
6-well Cell Culture Plate
9.5
2
12-well Cell Culture Plate
3.8
1
24-well Cell Culture Plate
1.9
0.6
96-well Cell Culture Plate
0.32
0.1
384-well Cell Culture Plate
0.06
0.025
T25 Flask
25
8
Table 1: Summary of Useful Volumes and Measures
All volumes and measures are
per well
, if applicable.
Note:
For glass coverslips for immunocytochemistry or electrophysiological
applications, see the iCell Cardiomyocytes Application Protocols available
online at www.cellulardynamics.com/lit/.
2.
Incubate the vessel(s) in a 37°C cell culture incubator for at least 1 hour.
3.
Aspirate gelatin solution immediately before addition of the cell suspension.
