Keros Victoria, Rosenlund Björn, Hultenby Kjell, Aghajanova Lusine, Levkov Lev, Hovatta Outi
Karolinska Institute, Division of Obstetrics and Gynaecology, Department of Clinical Science, Karolinska University Hospital, Huddinge, SE 141 86 Stockholm, Sweden.
Hum Reprod. 2005 Jun;20(6):1676-87. doi: 10.1093/humrep/deh797. Epub 2005 Apr 28.
Cryopreservation of testicular tissue is an option in fertility preservation for pre-pubertal boys who will lose spermatogenic cells as a result of chemotherapy. We compared three different protocols and cryoprotectants in cryopreservation of testicular tissue.
Testicular tissue obtained from 16 infertile men was evaluated by light microscopy(LM), immunostaining against MAGE-A4, transmission electron microscopy (TEM) and organ culture. Seminiferous tubules (1312) from non-frozen (n = 16) and frozen-thawed samples (n = 34) were studied following cryopreservation using protocols with either 1,2-propanediol (PrOH), glycerol or dimethylsulphoxide (DMSO) as cryoprotectants.
Normal structure was seen in 86 +/- 6% (mean +/- SD) of the fresh tissue. After freezing with DMSO, 70 +/- 6% and after PrOH, 37+/-3% of the tubules were judged to be good. When glycerol was used, the structure of the basal compartment of the tubules was severely damaged. The ultrastructure of the cryopreserved samples as revealed by TEM and MAGE-positive spermatogonia confirmed the findings. Cryopreserved Leydig cells maintained their morphology and ability to release testosterone in culture.
DMSO as a cryoprotectant (at a 0.7 mol/l concentration) proved to maintain the structure of testicular tissue, especially spermatogonia, after cryopreservation better than PrOH or glycerol.
对于因化疗将失去生精细胞的青春期前男孩，睾丸组织冷冻保存是一种生育力保存的选择。我们比较了三种不同的方案和冷冻保护剂在睾丸组织冷冻保存中的效果。
对16名不育男性获取的睾丸组织进行光学显微镜（LM）评估、MAGE - A4免疫染色、透射电子显微镜（TEM）检查和器官培养。使用1,2 - 丙二醇（PrOH）、甘油或二甲基亚砜（DMSO）作为冷冻保护剂的方案进行冷冻保存后，研究非冷冻（n = 16）和冻融样本（n = 34）中的生精小管（1312个）。
新鲜组织中86±6%（平均值±标准差）可见正常结构。用DMSO冷冻后，70±6%的小管被判定为良好；用PrOH冷冻后，37±3%的小管被判定为良好。使用甘油时，小管基底室的结构严重受损。TEM显示的冷冻保存样本的超微结构和MAGE阳性精原细胞证实了这些发现。冷冻保存的睾丸间质细胞在培养中保持其形态和释放睾酮的能力。
作为冷冻保护剂（浓度为0.7 mol/l）的DMSO在冷冻保存后比PrOH或甘油能更好地维持睾丸组织尤其是精原细胞的结构。
