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Basic protocol
|
| Washing and sterilization of pups with water and 70% ethanol, respectively |
0:10 |
The indicated time periods are for 5–7 rat pups. Timings can be scaled up and down according to the number of animals being used |
| Euthanasia of pups |
0:15 |
|
| Harvesting the hearts through dissection |
0:15 |
|
| Mincing the hearts into smaller pieces |
0:10 |
|
| Digestion of the cardiac tissue and collection of cells in horse serum |
1:30 |
|
| Seeding cells on culture dishes (Pre-plating) |
1:30 |
|
| Washing and resubstituting the cells with media (to remove unattached cardiomyocytes) |
0:10 |
|
|
Total
|
4:00 |
The total time may vary depending upon the number of animals being used for the experiment and the expertise of the user. |
|
Support protocol
|
|
|
| Adding poly-L-lysine coating solution and incubating |
0:30 |
|
| Removing the solution and keeping for air-drying |
0:30 |
The coating should be done either before or during the digestion of tissues. |
