Influence of different warming temperatures on the vitrified testicular fragments from pre‐pubertal cat

Abstract

Testicular vitrification is an alternative to preserve the genetic material of pre‐pubertal animals. However, there are few studies on post‐vitrification warming. Hence, the aim was to compare the influence of different warming temperatures on vitrified testicular fragments from pre‐pubertal cats. The testicles were fragmented and divided into a control group (non‐vitrified) and vitrified, using an association between dimethylsulphoxide and glycerol. The vitrified fragments were warmed at 50, 55 and 60°C/5 s. Morphological and morphometric evaluations were carried out using classical histology. Afterwards, the mitochondrial activity was evaluated using Rhodamine 123. The data were expressed in mean and standard error. The differences were considered significant when p < .05. In the histomorphological analysis, the testicular fragment presented seminiferous tubules with poorly developed germinal epithelium, compatible with pre‐pubertal animals. The group warmed at 50°C presented similar to the control regarding the maintenance of the integrity of the tubules and cells, without stromal rupture and lamina propria alteration, as well as regarding the maintenance of the junctions between the cells. The group warmed at 55°C showed reduction of the cell junctions, and the one warmed at 60°C had increased detachment of the basement membrane (p < .05). The warming caused a reduction in the tubular diameter inversely proportional and progressive to the increase in temperature, with the highest diameter in the control group and the lowest in the 60°C group. The control group showed a lower incidence of Rhodamine 123, followed in ascending order of the warmings at 55 and 60°C. The higher mitochondrial activity was obtained with 50°C, showing an increase of the metabolic cell function at this temperature. It was concluded that the testicular fragment of pre‐pubertal cats presents a better preserved morphology, morphometry and viability when warmed at 50°C.

Canine and Feline Testicular Preservation

“…In another work, three different warming temperatures (50, 55 and 60 °C for 5 s) were tested using the fresh testicular fragments as a control ( Figure 2 A). The aim of this work was to evaluate the possible influence of different warming temperatures on the structure, metabolic activity, composition, and cellular functions of the vitrified testicular fragments of prepubertal cats [ 27 ]. The authors concluded that vitrified testicular fragments from prepubertal cats have better preserved morphology, morphometry and viability when warmed at 50 °C ( Figure 2 B) compared to 60 °C ( Figure 2 C).…”