Ghavami Maryam, Mohammadnejad Daryoush, Beheshti Rahim, Solmani-Rad Jafar, Abedelahi Ali
Department of Anatomical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.
Drug Applied Research Center, Tabriz University of Medical Science, Tabriz, Iran.
J Reprod Infertil. 2015 Jul-Sep;16(3):138-47.
Cryopreservation of mammalian ovaries has been reported with different levels of success. Cryopreservation of ovarian tissue may be a potential alternative for treatment of infertility and many attempts have been done to improve the efficiency of ovarian cryopreservation. The objective of the present study was to compare the direct cover vitrification (DCV) with ethylene glycol (EG), dimethyl sulfoxide (DMSO) and EG plus DMSO.
Eighty five mice were sacrificed by cervical dislocation and their ovaries were cryopreserved in the presence of 5% EG or DMSO alone or as mixture, 10% EG or DMSO alone or as mixture and a group with ascending concentrations of cryoprotectants. After toxicity testing and vitrification warming, the ovaries were fixed for histological and ultrastructural studies. In addition, the viability of mechanically isolated follicles was studied by trypan blue staining. All data were compared by ANOVA (p<0.05).
Ovarian tissues frozen in EG plus DMSO in ascending concentrations retained a higher percentage of morphologically normal and or viable follicles than tissues frozen in 10 M EG plus DMSO or in either concentration of EG and DMSO alone (p<0.001). Ultrastructural analysis of ovarian tissues frozen in ascending concentrations of EG plus DMSO showed that these follicles were well preserved and it was very similar to the control group.
Cryopreservation of ovarian tissue in EG plus DMSO is the most effective method for preserving the structural integrity of follicles within the ovary.
已有关于哺乳动物卵巢冷冻保存取得不同程度成功的报道。卵巢组织冷冻保存可能是治疗不孕症的一种潜在替代方法，并且已经进行了许多尝试来提高卵巢冷冻保存的效率。本研究的目的是比较乙二醇（EG）、二甲基亚砜（DMSO）以及EG加DMSO的直接覆盖玻璃化法（DCV）。
通过颈椎脱臼法处死85只小鼠，将其卵巢在单独使用5%的EG或DMSO、或作为混合物使用，单独使用10%的EG或DMSO、或作为混合物使用以及一组含有浓度递增的冷冻保护剂的条件下进行冷冻保存。经过毒性测试和玻璃化复温后，将卵巢固定用于组织学和超微结构研究。此外，通过台盼蓝染色研究机械分离卵泡的活力。所有数据通过方差分析进行比较（p<0.05）。
与在10 M EG加DMSO中冷冻的组织或单独使用任何一种浓度的EG和DMSO冷冻的组织相比，在浓度递增的EG加DMSO中冷冻的卵巢组织保留了更高比例的形态正常和/或有活力的卵泡（p<0.001）。对在浓度递增的EG加DMSO中冷冻的卵巢组织进行超微结构分析表明，这些卵泡保存良好，与对照组非常相似。
在EG加DMSO中冷冻保存卵巢组织是保存卵巢内卵泡结构完整性的最有效方法。
