Antegrade Perfusion Model: An Ex Vivo Technique to Perfuse Isolated Mouse Heart
Antegrade Perfusion Model: An Ex Vivo Technique to Perfuse Isolated Mouse Heart
Transcription
Begin by placing an anesthetized mouse in the supine position. Dissect the thoracic cavity to visualize the heart. Use a pipettor fitted with a wide bore tip to apply suction pressure and aspirate the heart from the mouse’s body into the tip. Excise the heart at the aorta to harvest it from the mouse's body.
Immerse the isolated heart into a beaker containing a nutrient-rich buffer supplemented with an anticoagulant agent. The anticoagulant in the buffer forms a complex with calcium present in the cardiomyocytes – the heart muscle cells, ceasing contraction of cardiac muscles and preventing blood coagulation.
Next, place the heart apex-side down on a heart stand kept in a Petri dish containing suitable buffer. Clamp the aorta above the atria and transfer the clamped heart to a perfusion plate.
Subsequently, take a syringe-needle assembly containing an appropriate perfusion buffer. Insert the needle into the left ventricle piercing the ventricular chamber.
With the aorta clamped, intraventricular injection pressurizes aortic valves to open. Consequently, the buffer flushes blood antegradely through the coronary arteries, thoroughly perfusing heart muscles. Finally, the buffer effuses through the pulmonary veins. This provides a successfully perfused heart for downstream applications.