Effects of different oocyte retrieval and in vitro maturation systems on bovine embryo development and quality
Effects of different oocyte retrieval and in vitro maturation systems on bovine embryo development and quality
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DOI：
10.1017/s0967199413000658
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发表时间：
2015-06-01
期刊：
影响因子：
1.7
通讯作者：
Niemann, Heiner
中科院分区：
文献类型：
--
作者：
Bernal Ulloa, Sandra Milena;Heinzmann, Julia;Niemann, Heiner
环磷酸腺苷(CAMP)调节剂被用来避免卵母细胞的自发成熟，同时提高卵母细胞的发育能力。本工作评价了体外成熟过程中添加cAMP调节剂Forskolin、3-异丁基-1-甲基黄嘌呤(IBMX)和西洛胺对囊胚质量和产量的影响。实验分为三组：(I)切片或(Ii)在TCM199组织培养液中抽吸成熟24 h(分别为TCM24切片和TCM24抽吸)，(Iii)在cAMP调节剂存在下抽吸成熟30 h(cAMP30抽吸)和体内产生的囊胚。在体外成熟的卵母细胞受精，并在体外培养假定受精卵以评估胚胎发育。对卵裂率、囊胚形成、囊胚细胞数、选定基因的mRNA丰度和整体甲基化情况进行了评估。TCM24吸入法的囊胚率/受精率(32.2+/-2.1%)显著高于TCM24切片法和cAMP30吸入法(分别为23.4+/-1.2%和23.3+/-2.0%，P<0.05)，而其他组的囊胚率/受精率均显著高于TCM24切片法和cAMP30吸入法。结论：回收、收集系统和添加cAMP调节剂可以影响卵母细胞的发育能力，这不仅反映在囊胚率上，而且还反映在整体DNA甲基化和基因表达模式上。
Cyclic adenosine monophosphate (cAMP) modulators have been used to avoid spontaneous oocyte maturation and concomitantly improve oocyte developmental competence. The current work evaluated the effects of the addition of cAMP modulators forskolin, 3-isobutyl-1-methylxanthine (IBMX) and cilostamide during in vitro maturation on the quality and yields of blastocysts. The following experimental groups were evaluated: (i) slicing or (ii) aspiration and maturation in tissue culture medium (TCM)199 for 24 h (TCM24slicing and TCM24aspiration, respectively), (iii) aspiration and maturation in the presence of cAMP modulators for 30 h (cAMP30aspiration) and in vivo-produced blastocysts. In vitro-matured oocytes were fertilized and presumptive zygotes were cultured in vitro to assess embryo development. Cleavage, blastocyst formation, blastocyst cell number, mRNA abundance of selected genes and global methylation profiles were evaluated. Blastocyst rate/zygotes for the TCM24aspiration protocol was improved (32.2 +/- 2.1%) compared with TCM24slicing and cAMP30aspiration (23.4 +/- 1.2% and 23.3 +/- 2.0%, respectively, P 0.05), while those from the other groups were significantly elevated. It is concluded that retrieval, collection systems and addition of cAMP modulators can affect oocyte developmental competence, which is reflected not only in blastocyst rates but also in global DNA methylation and gene expression patterns.