Effect of time during transport of excised mare ovaries on oocyte recovery rate and quality after in vitro maturation
Effect of time during transport of excised mare ovaries on oocyte recovery rate and quality after in vitro maturation
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DOI：
10.1016/s0093-691x(99)00169-7
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发表时间：
1999-10-01
期刊：
影响因子：
2.8
通讯作者：
Palmer, E
中科院分区：
文献类型：
--
作者：
Guignot, F;Bezard, J;Palmer, E
在母马中，只有有限数量的卵母细胞可以成功地在体内收集，所以当需要大量的卵母细胞用于实验时，必须使用屠宰前收获的卵巢。从屠宰场到实验室处理和运输卵巢所产生的温度变化和时间间隔对卵母细胞的回收率及其在IVF和成熟后的质量产生不利影响。我们选择通过评估卵母细胞回收率、IVF前达到的核成熟阶段和IVF后达到的卵裂率来研究温度和运输时间对切除卵巢的影响，在短期（1.5至4小时）和长期（6至8小时）储存后。在短时间和长时间间隔内，储存容器中的温度分别从37 ℃降至32 ℃和27.5 ℃。卵丘-卵母细胞复合体（COC）被分类为具有致密的卵丘，完全或部分地围绕卵母细胞（致密的）;仅具有围绕卵母细胞的辐射冠（冠）的那些;具有完全或部分扩展的卵丘，显示围绕卵母细胞的细胞状或稀疏细胞状凝胶状云的那些（扩展的）;以及完全剥除卵丘和冠细胞的那些（剥除的）。所有的COCs，除了裸露的，这是丢弃的，在体外成熟30小时，在38.5摄氏度，在5%的CO2。长时间保存的卵母细胞回收率显著高于短时间保存的卵母细胞（48% vs 35%; P < 0.01），但采集的COCs在4类中的分布不受保存时间的影响。体外成熟后，卵母细胞核成熟度不受保存时间的影响，但保存时间短的卵母细胞胞质膜完整率高于保存时间长的卵母细胞（54% vs 34%; P = 0.07），保存时间长的卵母细胞胞质膜完整率高于保存时间长的卵母细胞（P < 0.01）。此外，在中期II（MII）与完整的膜的卵母细胞与短的存储间隔和冠COC类，而损坏的膜和不完全成熟与长期存储和紧凑的COC类。(C)1999年，Elsevier Science Inc.
In the mare only a limited number of oocytes can be successfully collected in vivo,so that when large numbers of oocytes are needed for experimentation, ovaries harvested front slaughtered mares must be used. The resulting temperature changes and time intervals mandated by handling and transport of ovaries from the slaughterhouse to the laboratory adversely affect the rate of oocyte recovery and their quality after IVF and maturation.We chose to study the effect of temperature and time in transit of excised ovaries by evaluating rate of oocyte recovery, nuclear maturation stage reached before, and cleavage rate reached after IVF, following short (1.5 to 4 h) and long (6 to 8 h) storage. Temperatures in the storage container decreased from 37-C to 32 degrees and 27.5 degrees C during the short and long interval, respectively. The cumulus-oocytes complexes (COCs) were classified as having a compact cumulus, completely or partially surrounding the oocyte (compact); those having only a corona radiata surrounding the oocyte (corona); those having a completely or partially expanded cumulus, showing a cellular or sparsely cellular, gelatinous cloud around the oocyte (expanded);and those that were completely denuded of both cumulus and corona cells (denuded). All COCs, except the denuded ones, which were discarded, were matured in vitro for 30 h at 38.5 degrees C in 5% CO2. The recovery rate of oocytes was significantly higher after long vs short storage (48 vs 35%; P < 0.01), but the distribution of the collected COCs into the 4 classes was not affected by the storage time. After in vitro maturation nuclear maturity was not affected by the storage time, but oocytes with intact cytoplasmic membranes were more frequently found after short than after long storage (54 vs 34%; P = 0.07), and fully matured oocytes were more often seen with intact membrane (P < 0.01). Moreover, oocytes with intact membranes in metaphase II (MII) were associated with short storage intervals and the corona COC class, while damaged membranes and incomplete maturation were associated with the long storage and the compact COC class. (C) 1999 by Elsevier Science Inc.