Cryopreservation of testicular tissue is a potentially effective method to preserve fertility in infertile patients who cannot obtain sperm or prepubertal boys who have cancer and require radiotherapy or chemotherapy. In this study

the vitrification of massive mouse testicular tissue was studied. Massive testicular tissue was immersed in vitrification solutions with different concentrations for different immersion time. Thermal analysis was conducted using a differential scanning calorimeter

and the testicular tissue was vitrified. The results showed that ice crystals were formed in the tissue during the cooling process in the low-concentration CPA group. In contrast

vitrification was achieved during the cooling process in the high-concentration CPA group. According to the negative rate of apoptosis of spermatogenic cells

the optimal vitrification loading protocol was to incubate in 20% DMSO for 1 min

followed by incubation with spermatogonial cells 78.6%

spermatoblast cells 90%

sperm cells 70.1%

and Sertoli cells 89.1%. Compared to slow freezing with vitrification

slow freezing can maintain the morphological integrity of testicular tissue and reduce the apoptotic rate of spermatogenic cells

making it more suitable for freezing massive testicular tissue.