Cardiopulmonary Complex Decellularization: A Technique for Decellularizing Heart and Lungs from Murine Model
Transcript
Decellularization devoids organs of their cellular components leaving the ECM structure intact. This helps in conserving the 3D architecture of organs. For the decellularization procedure, place a euthanized mouse in the supine position. Dissect the mouse and expose the cardiopulmonary complex.
Ligate all the major vessels originating from the heart except the aorta and major vessels of pulmonary circulation. This step prevents fluid leakage during the subsequent perfusion procedure. Create an opening in the trachea and insert a catheter through this opening. Catheterize the descending aorta from the bottom until the catheter reaches the aortic arch.
Suture the catheters to secure their position and connect them to the perfusion pump. Begin perfusion with deionized water. Perfusion through the aorta allows efficient removal of blood from the entire cardiac vasculature. Eventually, the water enters the pulmonary artery leading to the lungs.
After completely removing blood from the lung vasculature, stop water perfusion and replace it with different detergent solutions ranging from mild to strong. This procedure lyses cells releasing the intracellular components, making the heart and lungs appear translucent.
Perfuse again with deionized water to remove any remaining detergent molecules. Store decellularized cardiopulmonary complex in antibiotic-containing solution, which ensures a long shelf-life.
