Gook D A, Osborn S M, Johnston W I
Reproductive Biology Unit, Royal Women's Hospital, Carlton, Victoria, Australia.
Hum Reprod. 1993 Jul;8(7):1101-9. doi: 10.1093/oxfordjournals.humrep.a138201.
Human and mouse oocytes were cryopreserved by a slow freeze, rapid thaw method, using propanediol (PROH) as the cryoprotectant. A simulated cryopreservation was also included in the study to detect the level of damage attributable to the PROH alone. Comparison of the mouse and human oocytes cryopreserved by the same method showed opposing results, with a poor morphological survival rate of 4% observed for mouse oocytes and a subsequent normal fertilization rate of 0%. In 171 cryopreserved human oocytes a higher survival rate of 64% was achieved, and this showed more similarity to the mouse pronuclear oocytes survival of 53%. A comparison of human oocytes, cryopreserved within the cumulus and denuded of cumulus and corona prior to cryopreservation, demonstrated a higher survival rate in the denuded oocytes of 69% compared to 48%. A delay prior to cryopreservation of 1 or > or = 2 days had no effect on the immediate survival of oocytes, but culture for a further 24 h after thawing reduced survival, with the day 1 oocytes exhibiting the most dramatic reduction in survival (28%). Using a lectin binding method, abundant cortical granules were observed in all cryopreserved oocytes analysed. The meiotic spindle and chromosomes were examined in cryopreserved oocytes using fluorescence microscopy and 60% of the surviving oocytes had a normal spindle and chromosome configuration.
使用丙二醇(PROH)作为冷冻保护剂,通过慢速冷冻、快速解冻法对人和小鼠的卵母细胞进行冷冻保存。该研究还包括模拟冷冻保存,以检测仅由PROH引起的损伤程度。对采用相同方法冷冻保存的小鼠和人类卵母细胞进行比较,结果相反,小鼠卵母细胞的形态存活率低至4%,随后的正常受精率为0%。在171个冷冻保存的人类卵母细胞中,存活率较高,达到了64%,这与小鼠原核卵母细胞53%的存活率更为相似。对冷冻保存前在卵丘内以及去除卵丘和透明带的人类卵母细胞进行比较,结果显示去除卵丘的卵母细胞存活率更高,为69%,而有卵丘的为48%。冷冻保存前延迟1天或≥2天对卵母细胞的即时存活率没有影响,但解冻后再培养24小时会降低存活率,其中第1天的卵母细胞存活率下降最为显著(28%)。采用凝集素结合法,在所有分析的冷冻保存卵母细胞中均观察到丰富的皮质颗粒。使用荧光显微镜对冷冻保存的卵母细胞中的减数分裂纺锤体和染色体进行检查,60%存活的卵母细胞具有正常的纺锤体和染色体构型。