Serum maintains the fertilizability of mouse oocytes matured in vitro by preventing hardening of the zona pellucida
Serum maintains the fertilizability of mouse oocytes matured in vitro by preventing hardening of the zona pellucida
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血清通过防止透明带硬化来维持体外成熟的小鼠卵母细胞的受精能力
DOI：
10.1002/mrd.1120150203
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
J. Eppig
中科院分区：
文献类型：
--
作者：
S. M. Downs;A. Schroeder;J. Eppig
本文研究了卵母细胞体外成熟过程中血清和卵丘细胞对卵母细胞受精能力和透明质酸稳定性的影响。将卵丘细胞包封的卵母细胞在含有牛血清白蛋白和不同浓度的胎牛血清（FBS）的培养基中培养15 - 16小时。当血清浓度从5%逐渐降低至0%时，对糜蛋白酶消化的抗性以剂量依赖性方式相应增加。在无血清培养基中成熟的卵母细胞的透明质酸消化时间比在5%FBS中成熟的对照卵母细胞增加近500%。此外，随着血清浓度的降低，受精率降低;在无血清中成熟的卵母细胞的受精率降低了90%以上。这种受精能力的丧失与成熟卵子授精后卵黄内没有精子有关。将血清浓度从5%增加到10%对受精或透明带消化时间没有影响。血清剥夺，即使是很短的一段时间，在文化的发病显着降低卵丘细胞封闭的卵母细胞的受精能力，并增加了透明质酸消化时间。卵母细胞体外成熟后去除血清影响不大。在体外成熟过程中，完整卵丘的存在对于维持受精能力和胚胎稳定性是重要的。这些数据支持的想法，血清剥夺和/或去除卵丘细胞在卵母细胞体外成熟过程中的结果在改变透明质膜，表现为增加抵抗蛋白水解消化和精子穿透。
The effects of serum and cumulus cells during oocyte maturation in vitro on subsequent oocyte fertilizability and zona pellucida digestability have been examined. Cumulus cell-enclosed oocytes were cultured 15–16 hours in medium containing bovine serum albumin plus varying concentrations of fetal bovine serum (FBS). When the serum concentration was decreased incrementally from 5% to 0%, resistance to chymotrypsin digestion increased accordingly in a dose-dependent manner. The zona pellucida digestion time for oocytes matured in serum-free medium was increased nearly 500% above that for control oocytes matured in 5% FBS. Also, fertilization was decreased as serum concentration was lowered; the fertilization percentage for oocytes matured in serum-free serum was reduced by over 90%. This loss of fertilizability was correlated with an absence of sperm within the vitellus following insemination of matured ova. Increasing the serum concentration from 5% to 10% had no effect on fertilization or zona pellucida digestion time. Serum deprivation, even for a short period of time, at the onset of culture significantly reduced the fertilizability of cumulus cell-enclosed oocytes and increased the zona pellucida digestion time. Removal of serum after oocyte maturation in vitro had little effect. The presence of an intact cumulus oophorus during maturation in vitro was important in the maintenance of fertilizability and zona digestability. These data support the idea that serum deprivation and/or removal of the cumulus cells during oocyte maturation in vitro results in an alteration of the zona pellucida that is manifested as an increased resistance to proteolytic digestion and sperm penetration.