Lifegift Organ Donation Center 1 article published in JoVE Bioengineering Decellularization of Whole Human Heart Inside a Pressurized Pouch in an Inverted Orientation Doris A. Taylor1, Luiz C. Sampaio1, Rafael Cabello1, Abdelmotagaly Elgalad1, Rohan Parikh1, R Patrick Wood2, Kevin A. Myer2, Alvin T. Yeh3, Po-Feng Lee1 1Regenerative Medicine Research, Texas Heart Institute, 2Lifegift Organ Donation Center, 3Biomedical Engineering, Texas A&M University This method enables decellularization of a complex solid organ using a simple protocol based on osmotic shock and perfusion of ionic detergent with minimal organ matrix disruption. It comprises a novel decellularization technique for human hearts inside a pressurized pouch with real-time monitoring of flow dynamics and cellular debris outflow.