小鼠睾丸组织玻璃化保存的初步实验研究

对于无法取精的不育患者或患有癌症需放化疗的青春期前男孩，睾丸组织冷冻是潜在的有效保存生育力的方法。本文对小鼠块状睾丸组织的玻璃化保存方法进行了研究，运用差示扫描量热仪，对在不同体积分数保护剂中浸泡了不同时间后的小鼠块状睾丸组织进行热分析，随后对其进行玻璃化冻存，并将睾丸组织慢速冷冻保存与玻璃化冻存进行了对比。结果表明：低体积分数保护剂组在降温过程中检测到了组织内冰晶形成，而高体积分数保护剂组在降温过程中实现了玻璃化冷冻，未有冰晶形成；以生精细胞的凋亡阴性率为依据，筛选出体积分数为20%DMSO保护剂溶液中浸泡1 min后进行玻璃化冻存为最优玻璃化加载方案，其冻后各生殖细胞凋亡阴性率分别为精原细胞78.6%,精母细胞90%,精子细胞70.1%,支持细胞89.1%;与玻璃化冻存相比，慢速冷冻保存更能维持睾丸组织的冻后形态学完整性，并减小冻后生精细胞的凋亡率，是更适用块状睾丸组织冷冻的方法。

Experimental Study on Vitrification of Mouse Testicular Tissue

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.