Trehalose protects testicular tissue of dairy goat upon cryopreservation.

ABSTRACT

The aim of this study was to investigate whether the addition of trehalose to cryomedia reduces cellular damage and improves gene expression in cryopreserved dairy goat testicular tissues. Testicular tissues were cryopreserved in cryomedia without or with trehalose at a concentration of 5%, 10%, 15%, 20% or 25%. Cryopreserved testicular tissues were analysed for TUNEL-positive cell number, expression of BAX, BCL-2, CREM, BOULE and HSP70-2. Isolated Leydig cells from cryopreserved tissue were cultured, and spent medium was evaluated for testosterone level. The results showed that though the TUNEL-positive cell number increased in cryopreserved testicular tissues, the presence of trehalose reduced apoptotic cell number significantly. Quantitative real-time polymerase chain reaction results showed that although the expression of BAX was upregulated following cryopreservation, the presence of trehalose downregulates it in cryopreserved testicular tissues. Expression of BCL-2, CREM, BOULE and HSP70-2 was downregulated following cryopreservation but the presence of trehalose significantly upregulated their expression in cryopreserved testicular tissues. Leydig cells isolated from testicular tissues cryopreserved with trehalose produced higher testosterone than the one without it (control). These results suggest that trehalose has a protective role in cryopreservation of dairy goat testicular tissue, and the most suitable trehalose concentration for cryopreservation is 15%.