Pensis M, Loumaye E, Psalti I
Department of Obstetric and Gynecology, University of Louvain, Brussels, Belgium.
Fertil Steril. 1989 Nov;52(5):787-94. doi: 10.1016/s0015-0282(16)61032-4.
One hundred and twenty-one freshly-collected human oocytes and 839 unfertilized human oocytes after insemination were cryopreserved by vitrification. The cryoprotectants used were dimethylsulfoxide (DMSO) and sucrose. Vital staining and morphological criteria were used to assess injuries to cells. Variation of the time exposure to DMSO and sucrose, and cryoprotectants concentrations, followed by extraction-dilution in sucrose without freezing made it possible to study chemical toxicities. Variation of cryoprotectant concentrations followed by immersion in liquid nitrogen, thawing, extraction, and dilution made it possible to choose optimal conditions for vitrification. The sucrose concentration upon extraction after freezing and thawing which was lower than that during soaking enhanced the oocyte survival rate as did the choice of duration and temperature of soaking. No parthenogenetical activation of these unfertilized ovum was observed. This study indicates that with a certain combination of DMSO and sucrose concentrations up to 80% of morphologically intact human oocytes can be recovered after rapid freezing and thawing.
121枚新鲜采集的人类卵母细胞和839枚授精后未受精的人类卵母细胞通过玻璃化法进行冷冻保存。所使用的冷冻保护剂为二甲基亚砜（DMSO）和蔗糖。采用活细胞染色和形态学标准评估细胞损伤情况。改变DMSO和蔗糖的暴露时间以及冷冻保护剂浓度，随后在不冷冻的情况下于蔗糖中进行提取 - 稀释，从而能够研究化学毒性。改变冷冻保护剂浓度，随后进行液氮浸泡、解冻、提取和稀释，从而能够选择玻璃化的最佳条件。冻融后提取时的蔗糖浓度低于浸泡时的蔗糖浓度，这与浸泡时间和温度的选择一样，提高了卵母细胞的存活率。未观察到这些未受精卵母细胞的孤雌激活现象。本研究表明，通过DMSO和蔗糖浓度的特定组合，快速冷冻和解冻后高达80%的形态完整的人类卵母细胞能够得以恢复。
