Developmental competence and gene expression of immature oocytes following liquid helium vitrification in bovine
Developmental competence and gene expression of immature oocytes following liquid helium vitrification in bovine
复制标题
牛液氦玻璃化冷冻后未成熟卵母细胞的发育能力和基因表达。
DOI：
10.1016/j.cryobiol.2014.09.380
复制
发表时间：
2014-12-01
期刊：
影响因子：
2.7
通讯作者：
Yu, Xue-Li
中科院分区：
文献类型：
--
作者：
Chen, Jun-Yi;Li, Xiao-Xia;Yu, Xue-Li
本研究旨在建立一种有效的超快速玻璃化冷冻方法，并评价其对牛未成熟卵母细胞成熟、发育能力及发育相关基因表达的影响。将牛卵丘卵母细胞复合体随机分为3组：（1）对照组，（2）液氮玻璃化冷冻组，（3）液氦玻璃化冷冻组。将卵母细胞玻璃化冷冻后温热，液氦组形态正常的卵母细胞比例（89.0%）显著高于液氮组（81.1%）（P < 0.05）。玻璃化冷冻解冻卵母细胞体外成熟24 h后，液氦组的成熟率（50.6%）显著高于液氮组（42.6%）（P < 0.05）。体外受精后，液氦玻璃化冷冻组的卵裂率和囊胚率（41.1%和10.0%）显著高于液氮玻璃化冷冻组（33.0%和4.5%，P < 0.05）。采用实时荧光定量聚合酶链反应（qRT-PCR）检测玻璃化冷冻-解冻卵母细胞成熟24 h后GDF 9、BAX和ZAR 1的表达。这些基因的表达在玻璃化冷冻后发生了改变。GDF 9和BAX在液氦玻璃化冷冻组与对照组之间的表达差异无统计学意义，而在液氮玻璃化冷冻组与对照组之间的表达差异有统计学意义。因此，液氦玻璃化冷冻牛未成熟卵母细胞是可行的。玻璃化卵母细胞的发育能力受损与这些基因表达水平的改变之间存在联系。(C)2014爱思唯尔公司All rights reserved.
The objective of this study was to develop an effective ultra-rapid vitrification method and evaluate its effect on maturation, developmental competence and development-related gene expression in bovine immature oocytes. Bovine cumulus oocyte complexes were randomly allocated into three groups: (1) controls, (2) liquid nitrogen vitrification, and (3) liquid helium vitrification. Oocytes were vitrified and then warmed, the percentage of morphologically normal oocytes in liquid helium group (89.0%) was significantly higher (P < 0.05) than that of the liquid nitrogen group (81.1%). When the vitrified-thawed oocytes were matured in vitro for 24 h, the maturation rate in liquid helium group (50.6%) was higher (P < 0.05) than liquid nitrogen group (42.6%). Oocytes of liquid helium vitrification had higher cleavage and blastocyst rates (41.1% and 10.0%) than that of liquid nitrogen vitrification (33.0% and 4.5%; P < 0.05) after in vitro fertilization. Moreover, the expression of GDF9 (growth/differentiation factor-9), BAX (apoptosis factor) and ZAR1 (zygote arrest 1) was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) when the vitrified-thawed oocytes were matured 24 h. The expression of these genes was altered after vitrification. Expression of GDF9 and BAX in the liquid helium vitrification group was not significantly different from that of the control, however there were significant differences between the liquid nitrogen vitrification group and control. In conclusion, it was feasible to use liquid helium for vitrifying bovine immature oocytes. There existed an association between the compromised developmental competence and the altered expression levels of these genes for the vitrified oocytes. (C) 2014 Elsevier Inc. All rights reserved.