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Ensure system is primed.
Watch Power Point of how to Prime System
When cannulating the aorta ensure flow rate is sufficient to sustain the heart of the species being used.
Mouse 3 – 5 mL/min
Rat and guinea pig 5 – 10 mL/min
Rabbit 20 – 30 mL/min.
Maintain flow while removing the heart of the donor animal.
Heart is quickly excised from the donor animal and is placed in ice cold buffer solution (Krebs-Henseleit, Tyrodes, etc.)
It is better to have more aorta than less aorta
Lungs may or may not be removed with the heart. An added benefit of leaving the lungs attached to the heart is the heart will “float” in the ice-cold buffer making retrieval easier.
Prepare too mount the heart.
Quickly locte the aorta. Look for the vessel that conspicuously spurts blood.
If the lungs were removed attached to the heart, the aorta will be located between the left and right lobes of the lungs.
Using forceps claspthe edges of the open end of the aorta and slip it on the aortic cannula.
Ensure that the bottom edge of the cannula does not extend into the aortic valves. If the aorta is placed too far up the cannula, damage to the aortic valves may occur.
Mounting the heart.
Clamp the aorta to the cannula using a bulldog clamp and tie the aorta to the cannula. For larger rabbit hearts, a modified electrical alligator clip can be used.
Adjust the heart/aortic position on the cannula so that flow is not out a branch off of the aortic arch but is going into the coronary ostia above the aortic valves. Coronary perfusion pressure should be your guide to proper positioning of the cannula. Position the heart so that the left and right atria are facing you.
Remove lungs and pericardium.
Quickly but carefully remove the lungs and pericardium.At this point it is better to remove too little tissue than too much. Ensure that all lung tissue is removed. It is best to slowly tease away tissue and cut small “thin” sections off than to remove large amounts of tissue when looking for the left atrial opening. You can remove more tissue if needed but you cannot add tissue already removed.
Suture aorta to cannula.
Tie the aorta to the cannula using a large diameter suture, no smaller than 4-0. Smaller sutures may cut through the aorta
The heart is mounted.
The Heart is now mounted and can be lowered into the Radnoti Heart Chamber.
Time from removal of the heart from the donor animal to initiating Langendorff flow should be less than 90 seconds. If the time to do this is greater than 90 seconds the heart may be compromised.
