Improved cryotolerance and developmental competence of human oocytes matured in vitro by transient hydrostatic pressure treatment prior to vitrification
Improved cryotolerance and developmental competence of human oocytes matured in vitro by transient hydrostatic pressure treatment prior to vitrification
复制标题
玻璃化冷冻前瞬时静水压处理可改善体外成熟的人类卵母细胞的低温耐受性和发育能力
DOI：
10.1016/j.cryobiol.2016.12.009
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发表时间：
2017-04-01
期刊：
影响因子：
2.7
通讯作者：
Sun, Yijuan
中科院分区：
文献类型：
--
作者：
Gu, Ruihuan;Feng, Yun;Sun, Yijuan
背景：目前，中期II期(MII)卵母细胞具有非常特殊的结构，导致其玻璃化冷冻存在复杂的困难，其效果仍需大量研究观察。本研究旨在探讨玻璃化冷冻前对卵母细胞施加瞬时静水压力(THP)对卵母细胞的发育能力和活性氧的影响，对卵母细胞的减数分裂纺锤体、透明带和DNA拷贝数无明显影响。方法：本研究的未成熟卵均于2015年2月至2015年12月在上海吉艾遗传与体外受精研究所采集。将MII期卵母细胞随机分为3组：新鲜卵母细胞组、玻璃化冷冻组和THP处理后玻璃化冷冻组。卵胞浆内单精子注射和胚胎培养后评估胚胎发育结局。此外，对3组受精卵的纺锤体行为、活性氧(ROS)、透明带(ZP)和DNA拷贝数变异进行了评估和比较。结果：共纳入568个MII卵母细胞。第3天B组胚胎细胞数明显少于A组和C组(A组5.01+/-2.11，B组3.89+/-2.21，C组4.69+/-2.05)。A组和C组的囊胚发育状况均优于B组。MII期卵母细胞在玻璃化冷冻前进行了THP处理，玻璃化冷冻的平衡时间(244.9±30.1vs.181.5+/-10.1)明显缩短。C组玻璃化冷冻组在玻璃化冷冻前用THP改善卵母细胞的ROS、ZP。THP对玻璃化冷冻/冷冻卵母细胞的减数分裂纺锤体和DNA拷贝数变异无影响。结论：玻璃化冷冻前亚致死性THP处理提高了人体外成熟卵母细胞的发育能力，减少了卵母细胞玻璃化冷冻过程中与玻璃化有关的ROS变化，对卵母细胞减数分裂纺锤体、ZP和DNA拷贝数变异无明显影响。(C)2017 Elsevier Inc.保留所有权利。
Background: At present, the metaphase II (MII) oocytes have a very special structure that leads to complex difficulties associated with its vitrification, and their efficacy still needs a large amount of study to observe. The present study was to investigate whether transient hydrostatic pressure (THP), which was utilized for oocytes before vitrification, had positive effect on the oocytes' developmental ability and reactive oxygen species, and had no damage on meiotic spindle, zona pellucida, and DNA copy number.Methods: All the immature oocytes used in this research were collected between February 2015 and December 2015 in Shanghai Ji Ai Genetics & IVF Institute. The MII oocytes, which were originated from metaphase I (MI) oocytes, were randomly distributed into three groups: A) fresh oocytes; B) vitrification; and C) vitrification after THP treatment. The embryo developmental outcome was evaluated after intracytoplasmic sperm injection and embryo culture. Furthermore, the meiotic spindle behavior, reactive oxygen species (ROS), zona pellucida (ZP), and DNA copy number variation were evaluated and compared among the three groups.Results: A total of 568 MII oocytes were included in the study. Embryos from group B had fewer cells on day 3 compared with group A and C (5.01 +/- 2.11 for group A, 3.89 +/- 2.21 for group B, and 4.69 +/- 2.05 for group C). The developmental feature of blastocyst in groups A and C were superior to those of group B. The MII oocytes were manipulated with THP before vitrification, and the equilibration time was significantly shorter in the vitrification procedure (244.9 +/- 30.1 vs. 181.5 +/- 10.1). The ROS, ZP of vitrified/warmed oocytes in group C were improved with THP before vitrification. THP had no influence on the meiotic spindle and DNA copy number variation of vitrified/warmed oocytes.Conclusions: The results of the study indicated that sublethal THP treatment before vitrification increased the developmental competence of human in vitro matured oocytes, reduced vitrification related changes in the ROS, which occurred during oocyte vitrification, and did not damage the meiotic spindle, ZP and DNA copy number variation. (C) 2017 Elsevier Inc. All rights reserved.