EA 4308 Spermatogenesis and Male Gamete Quality, Reproductive Biology Laboratory-CECOS, Rouen University Hospital, Institute for Biomedical Research, University of Rouen, Rouen, France.
Theriogenology. 2011 Oct 1;76(6):981-90. doi: 10.1016/j.theriogenology.2011.04.025. Epub 2011 Jun 12.
Fertility preservation has been included in the management of childhood cancer treatment. Cryopreservation of immature testicular tissue is the only available solution for pre-pubertal boys. Different freezing protocols have been developed in several species but without a clearly identified procedure. We tried to evaluate several protocols for cryopreservation of rat immature testicular tissue. Twelve different freezing protocols using different (i) cryoprotectant (dimethylsulphoxide [DMSO] or 1,2-propanediol [PROH]), (ii) cryoprotectant concentration (1.5M or 3M), (iii) equilibration time (30 or 60 min), (iv) equilibration temperature (4 °C or room temperature), (v) size of testicular fragment (7.5mg or 15 mg), (vi) package (straws or cryovials), were compared using cord morphological damage evaluation. A testicular tissue piece of 7.5mg cryopreserved in cryovial using 1.5M DMSO, an equilibration time of 30 min at 4 °C showed fewer morphological alterations than the other protocols tested. The selected freezing protocol was able to maintain rat immature testicular tissue architecture, functionality after testicular pieces organotypic culture, and could be proposed in a human application.
生育力保存已纳入儿童癌症治疗管理。不成熟睾丸组织的冷冻保存是青春期前男孩唯一可用的解决方案。已经在几种物种中开发了不同的冷冻保存方案，但没有明确的程序。我们试图评估几种大鼠未成熟睾丸组织冷冻保存方案。使用不同的（i）冷冻保护剂（二甲亚砜[DMSO]或 1,2-丙二醇[PROH]）、（ii）冷冻保护剂浓度（1.5M 或 3M）、（iii）平衡时间（30 或 60 分钟）、（iv）平衡温度（4°C 或室温）、（v）睾丸组织片段大小（7.5mg 或 15mg）、（vi）包装（细管或 cryovials），对 12 种不同的冷冻方案进行了比较，评估了脐带形态损伤。使用 1.5M DMSO 在 cryovial 中冷冻保存 7.5mg 睾丸组织，在 4°C 下平衡 30 分钟的方案显示出比其他测试方案更少的形态改变。所选的冷冻保存方案能够维持大鼠未成熟睾丸组织的结构和功能，在睾丸组织器官培养后，可在人类应用中提出。
