Vitrification of In vitro-matured Oocytes: Effects of meiotic spindle morphology on clinical outcome
Vitrification of In vitro-matured Oocytes: Effects of meiotic spindle morphology on clinical outcome
复制标题
体外成熟卵母细胞的玻璃化冷冻：减数分裂纺锤体形态对临床结果的影响
DOI：
10.4103/2096-2924.281854
复制
发表时间：
2020-01-01
影响因子：
0.8
通讯作者：
Sun, Yi-Juan
中科院分区：
文献类型：
--
作者：
Gu, Rui-Huan;Li, Zhi-Chao;Sun, Yi-Juan
目的：减数分裂纺锤体控制染色体的运动，并介导受精和受精后早期事件所必需的各种功能。本研究旨在研究玻璃化冷冻是否会导致体外成熟中期II（MII）人类卵母细胞减数分裂损伤，以及减数分裂纺锤体形态是否会影响随后的发育结果。研究方法：应用PolScope成像技术和免疫荧光染色技术对人MII期卵母细胞玻璃化冷冻前后的纺锤体特征进行了研究。并对卵母细胞的发育能力进行了检测。结果：在我院进行的593个卵胞浆内单精子注射周期中，共获得419个人类MII期卵母细胞。在这些卵母细胞中，54个用于免疫荧光染色，而其他卵母细胞通过PolScope成像进行检查，并根据减数分裂纺锤体形态分为三组：（A）正常形态，（B）弱折射和短减数分裂纺锤体，和（C）未检测到减数分裂纺锤体。三组在玻璃化冷冻后的存活率方面表现出统计学显著差异。然而，在玻璃化冷冻前后进行的免疫荧光染色中，未发现卵母细胞染色体结构和减数分裂纺锤体形态的差异。A组的卵母细胞存活率、受精率和早期胚胎发育率显著高于B组和C组（有或无玻璃化冷冻）。虽然玻璃化对A组中的这些指标没有影响，但B和C组在玻璃化/升温后表现出显著较低的受精率和卵裂率。结论：在玻璃化冷冻前筛选正常的减数分裂纺锤体形态和染色体构型可能会增加用于各种辅助生殖技术的健康活卵母细胞的产量。
Objective: The meiotic spindle controls chromosome movement and mediates various functions essential for fertilization and early postfertilization events. This study aimed to examine whether vitrification causes meiotic damage in vitro- matured metaphase II (MII) human oocytes, and whether the meiotic spindle morphology influences the subsequent developmental outcomes. Methods: The spindle characteristics of MII human oocytesin vitro matured were studied before and after vitrification using PolScope imaging and immunofluorescence staining. The developmental competence of oocytes was also examined. Results: A total of 419 human MII oocytes were obtained from 593 intracytoplasmic sperm injection cycles at our hospital. Of these oocytes, 54 were used for immunofluorescence staining, whereas the other oocytes were examined by PolScope imaging and classified into three groups according to the meiotic spindle morphology: (A) normal morphology, (B) weak refraction and short meiotic spindle, and (C) no detectable meiotic spindle. The three groups demonstrated statistically significant differences in terms of survival after vitrification. However, differences were not found in terms of oocyte chromosome structure and meiotic spindle morphology on immunofluorescence staining performed before and after vitrification. Oocyte survival, fertilization, and early embryonic development rates were significantly higher in Group A than in Groups B and C with or without vitrification. While vitrification had no effect on these metrics in Group A, Groups B and C demonstrated significantly lower fertilization and cleavage rates after vitrification/warming. Conclusions: Screening for normal meiotic spindle morphology and chromosome configuration before vitrification may increase the yield of healthy viable oocytes for various assisted reproductive technologies.