Abstract 15288: Effect of a Novel ORCA Bioreactor on Intact Porcine Heart Decellularization
Objectives: Heart disease causes significant mortality worldwide, making regeneration of cardiac tissue a pivotal goal. Using a decellularized heart as a biological scaffold for organogenesis provides a future application for clinical cardiac transplantation. We developed a decellularization protocol for porcine heart utilizing a novel organ regenerative control acquisition (ORCA) bioreactor.
METHODS: The ORCA bioreactor system consists of a temperature and pressure regulated monitor that simulates physiological conditions. The system was modified by gradual flow rates (≤2200ml/min) and pressures (≤400 mmHg) to decellularize porcine hearts (n=6). An infrared camera was used to measure surficial temperature and organ homogeneity throughout the process. Decellularization involved perfusion with 1-3% triton and 1-2% sodium dodecyl sulfate. Native and decellularized hearts were analyzed via DNA assessment, MRI imaging, histology and transmission electron microscopy (TEM).
RESULTS: Compared to a native heart, a heart decellularized in less than 10 hours appeared translucent, indicating loss of cellular components. Quantitative DNA analysis showed a significant decrease of DNA concentration compared to the control. Infrared images confirmed a homogeneous decellularization. MRI imaging displays retention of gross anatomy, vascularity and collagen bundles. Histology and TEM data indicated removal of nuclear material with the maintenance of structural architecture.
CONCLUSION: Cardiac decellularization requires pressures greater than those physiologically required. Gradual increases in flow rates and pressures may play a key role in decellularization and provide a viable scaffold for re-seeding with stem cells. Monitoring the decellularization protocol via ORCA system has resulted in a shorter cardiac decellularization method. This significantly impacts tissue regeneration by providing an improved model with clinical translation.
- © 2013 by American Heart Association, Inc.