Procurement and preservation of neonatal porcine cardiac tissue

The porcine and human heart are remarkably similar in cardiac physiology and biochemistry. Translational research involving the porcine biomedical model is becoming increasingly applicable for the study of human cardiac function in health and disease. Presently, few protocols exist for collecting experimentally viable cardiac tissue from large animal models, particularly during neonatal maturation. To address this deficiency, we have developed a technique to procure and preserve ventricular tissue from neonatal piglets at day 3 (n = 4) and day 30 (n = 6) postpartum. Piglets were subjected to a strict sedation, anesthesia, and analgesia regimen. During surgery, cardiopulmonary indices of electrocardiogram, heart rate, systolic and diastolic blood pressure, respiration rate, peripheral O₂ saturation, and end-tidal CO₂ were monitored continuously to ensure normal cardiac function. Before cardiectomy, each heart was perfused with an intravenous administration of heparin (10 mL/kg) and ice-cold Custodiol HTK cardioplegia solution (10 mL/kg). After cardiac explantation, myocardial samples (dimensions: 1 × 1 × 1 cm) were dissected from the left and right ventricles and snap-frozen in liquid nitrogen. Analysis via SDS-PAGE densitometry demonstrated that myofibrillar proteins are stable and undegraded. Western Blots showed full expression of protein. These results suggest that the detailed cardiac tissue procurement technique preserves the experimental viability of the cardiac tissue and prevents the degradation of myofibrillar proteins.NEW & NOTEWORTHY This project's objective was to develop a technique for procuring and preserving cardiac tissue from a porcine model. Porcine is a rapidly emerging animal model to study cardiovascular disease and has recently been popularized due to advancements in CRISPR-cas9 technology. The technique, originally derived from human heart transplant protocols, involves full anesthetic and analgesic regimens and the use of saline heparin and cardioplegia solution to preserve tissue integrity.