Refining the Guinea Pig (Cavia porcellus) Cryo-Injury Model for Cardiac Regeneration and Functional Characterization

Guinea pigs have been a standard model in cardiovascular pharmacology and physiology research, but the advent of transgenic models has largely replaced them with mouse and rat models. However, guinea pigs remain important models in cardiac electrophysiology, drug-induced arrhythmias, or atherosclerosis research, and they have recently gained importance for studying one specific research question, that is, transplantation of pluripotent stem cell derived cardiomyocytes to repair the cryo-injured heart. Their human-like cardiac electrophysiology, together with their small size that facilitates handling and housing, make guinea pigs a valuable experimental model for these studies. However, repeated open heart surgeries in guinea pigs are technically demanding and accompanied by high mortality. In this study, we retrospectively examined sequential protocol modifications and describe how protocol refinements led to improved survival rates. Cryo-injury was performed in female Dunkin-Hartley guinea pigs under general anesthesia with a liquid nitrogen-cooled probe via a lateral thoracotomy. Cells were transplanted during a second surgery 7 days later. We analyzed data from up to 558 animals to determine mortality rates and morphologic and functional parameters. Initial studies revealed a mortality rate of ∼50%. Sequential modifications led to a significant reduction, with the refined protocol achieving a perioperative mortality rate of ∼30%. The procedures were completed in <35 minutes, and survival rates for the observation period (up to 8 weeks) were 70%. Scar size was evaluated in 144 (4 weeks, n = 92; 8 weeks, n = 52) animals and showed a significant, but shallow correlation with echocardiographically determined heart function. Taken together, refined surgery protocols allow safe and reproducible cryo-injury with subsequent cell injections in guinea pigs with an improved mortality rate.