Understanding critical roles of warming and reanimation is critical to improve the survival of vitrified testicular tissue in domestic cats. The objective was to study structural and functional properties of testicular tissues from prepubertal domestic cats after standard vitrification followed by two warming protocols (directly at 37°C or with a 5-second pre-exposure to 50°C) and three reanimation time points (immediately, 24 h and 5 days post-warming). In Experiment 1, tissues were evaluated for histo-morphology and mitochondrial activity immediately or 24 h after warming protocols. In Experiment 2, cell viability, DNA fragmentation, and germ cell composition were assessed immediately, 24 h, or 5 days after optimal warming. Preservation of seminiferous tubule structure was better using warming at 50°C for five seconds, and survival of somatic as well as germinal cells was higher compared to direct warming at 37°C for one minute. Short term in vitro culture (for reanimation) also proved that cellular composition and functionality were better preserved when warmed for a short time at 50°C. Collective data showed that short warming at 50°C led to better quality of seminiferous tubule structure and cell composition after vitrification and short-term culture. In addition, data suggest clear directions to further understand and optimize testicular tissue survival after fertility preservation procedures.

Measurement of testicular artery blood flow is used in several species to evaluate reproductive function and testicular and scrotal pathology. In dogs there are inconsistent reports about normal flow in post-pubertal dogs and no information concerning pre-pubertal dogs. The aim of this study was to describe regional differences in testicular artery blood flow in clinically normal post-pubertal and pre-pubertal dogs with no history of reproductive tract disease. The post-pubertal dogs produced normal ejaculates throughout the study. In all dogs the three different regions of the artery were imaged and monophasic flow with an obvious systolic peak and flow throughout diastole was observed on every occasion. The highest peak systolic velocity (PSV) and end diastolic velocity (EDV) were measured within the distal supra-testicular artery and marginal artery whilst the lowest PSV and EDV were measured within the intra-testicular arteries. Flow measurements were not different between left and right testes and were consistent between dogs on different examination days. Calculated resistance index (RI) and pulsatility index (PI) were lowest in the intra-testicular arteries. The pre-pubertal dogs had significantly smaller testes than the post-pubertal dogs (p < 0.05) and were unable to ejaculate during the study. The three different artery regions were imaged at every examination time point, and flow profiles had a similar appearance to those of the post-pubertal dogs. PSV, EDV, RI and PI showed a similar trend to the post-pubertal dogs in that values were lowest in the intra-testicular arteries. Notably, values of PSV, EDV, RI and PI were significantly lower (p < 0.05) in pre-pubertal dogs compared with post-pubertal dogs. This study demonstrated important regional and pubertal differences in testicular artery blood flow of dogs, and form the basis for establishing baseline reference values that may be employed for the purposes of clinical diagnosis.