Figure 4:
CPAs cocktail 6 partially retained cardiac function of adult zebrafish hearts after 24 h, 72 h and 120 h of storage at −10 °C when loaded via perfusion (n = 6 animals per treatment). (A) Heart beat analysis for hearts stored in CPAs 6 resulted in similar heart rate recovery at all storage times. No statistical difference was seen between heart rate of CPAs cocktail 6 hearts over time or when compared to the beat ratio of hearts stored in clinical standard conditions (HypothermicCON) at each individual storage times. (B) Similarly, storage in CPAs cocktail 6 resulted in ~40% of cardiac tissue to remain viable post-thaw, irrespective of freezing time, suggesting temperature dependent injury but no time dependent injury. (C) Separate analysis of ventricular and atrial beat for hearts stored in CPAs 6 resulted in similar atrial and ventricular contraction (contraction per minute – cpm) recovery at all storage times, with the atrium contracting over twice as often as the ventricle. No statistical difference was seen between beat ratio of CPAs cocktail 6 hearts over time or when compared to the beat ratio of hearts stored in clinical standard conditions (HypothermicCON) at each individual storage times (D) Representative histological images of TUNEL and DAPI staining. Scale bar 250 μm. All statistical data is expressed as mean ± standard deviation. Heart rate and Cardiac Viability were analyzed via a two-way ANOVA and Tukey-Kramer HSD post-hoc analyses. Contraction rate was analyzed via an ordinary one-way ANOVA with Tukey’s multiple comparison test. **** = p < 0.000