Canine IVM With SOF Medium, Insulin-Transferrin-Selenium, and Low O(2) Tension Improves Oocyte Meiotic Competence and Decreases Reactive Oxygen Species Levels.
Canine IVM With SOF Medium, Insulin-Transferrin-Selenium, and Low O(2) Tension Improves Oocyte Meiotic Competence and Decreases Reactive Oxygen Species Levels.
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DOI：
10.3389/fcell.2021.694889
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发表时间：
2021
影响因子：
5.5
通讯作者：
Salamone DF
中科院分区：
文献类型：
--
作者：
Duque Rodriguez M;Cittadini CO;Teplitz GM;De Stefano A;Lombardo DM;Salamone DF
由于体外成熟（IVM）效率低，犬种的辅助生殖技术受到限制。与其他哺乳动物不同，母犬在生殖囊期排卵卵母细胞，并在输卵管环境中48-72小时后完成中期II（MII）并可受精。为此，我们比较了两种不同的IVM培养基，补充有8%牛血清白蛋白（BSA）的合成输卵管液（SOF）或8% BSA-2.5%胎牛血清（FBS）和TCM-199与10% FBS的混合物。此外，我们评估了补充胰岛素-转铁蛋白-硒（ITS）和低O2张力对卵母细胞成熟、活性氧（ROS）水平、膜完整性和孤雌激活（PA）后胚胎发育的影响。培养72小时后，SOF + BSA、SOF + BSA + FBS和TCM-199 + FBS分别显示5%、7%和4%的MII，无统计学差异。然而，与SOF + BSA + FBS相比，SOF + BSA产生显著更高的变性率（分别为44%和23%）。值得注意的是，与其他组相比，在高O2压力下补充1 μl/ml的ITS通过将成熟率提高高达20%而表现出有益效果。低O2张力的成熟率增加到36.5%，虽然没有统计学差异相比，高O2张力的ITS的存在。较低的ROS水平和较高的完整性的细胞质膜被发现在ITS的存在下，尽管没有差异，在低O2张力组的成熟率。此外，PA后，1%的发展，直到八细胞阶段后，在ITS的存在下激活体外成熟的卵母细胞。这些结果表明，添加8%BSA和2.5%FBS的SOF适合犬卵母细胞体外成熟，添加ITS对高、低O2压均有利。此外，在培养系统中添加ITS降低了IVM后家犬卵母细胞中的ROS水平并增加了膜完整性。
Assisted reproductive technologies in canine species are limited due to the low efficiency of in vitro maturation (IVM). Unlike other mammals, bitches ovulate oocytes in the germinal vesicle stage and complete metaphase II (MII) after 48–72 h in the oviductal environment and become fertilizable. For this reason, we compared two different IVM media, synthetic oviductal fluid (SOF) supplemented with 8% bovine serum albumin (BSA) or a mixture of 8% BSA–2.5% fetal bovine serum (FBS) and TCM-199 with 10% FBS. Additionally, we evaluated the effect of supplementation with insulin-transferrin-selenium (ITS) and low O2 tension in oocyte maturation, reactive oxygen species (ROS) levels, membrane integrity, and embryo development following parthenogenetic activation (PA). After 72 h of culture, SOF + BSA, SOF + BSA + FBS, and TCM-199 + FBS show 5, 7, and 4% of MII, respectively, without a statistical difference. However, SOF + BSA produced significantly higher degeneration rates compared to SOF + BSA + FBS (44 and 23%, respectively). Remarkably, supplementation with 1 μl/ml of ITS under high O2 tension demonstrated a beneficial effect by improving maturation rates up to 20% compared to the other groups. Low O2 tension increased maturation rates to 36.5%, although there were no statistical differences compared to high O2 tension in the presence of ITS. Lower ROS levels and higher integrity of the cytoplasmic membrane were found in the presence of ITS despite no differences in maturation rates under low O2 tension groups. Additionally, after PA, 1% development until the eight-cell stage was obtained after activation of in vitro-matured oocytes in the presence of ITS. Taken together, these results indicate that SOF supplemented with 8% BSA and 2.5% FBS is suitable for IVM of canine oocytes and ITS supplementation was beneficial for both high and low O2 tension. Furthermore, the addition of ITS in the cultured system lowers ROS levels and increases membrane integrity in domestic dog oocytes after IVM.
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影响因子：
3.7
作者：
Buemo CP;Gambini A;Moro LN;Hiriart MI;Fernández-Martín R;Collas P;Salamone DF
通讯作者：
Salamone DF
影响因子：
2.8
作者：
Cordova, B.;Morato, R.;Mogas, T.
通讯作者：
Mogas, T.
影响因子：
2.8
作者：
Luvoni, Gaia Cecilia
通讯作者：
Luvoni, Gaia Cecilia
影响因子：
2.8
作者：
Luvoni, GC;Chigioni, S;Macis, D
通讯作者：
Macis, D
影响因子：
2.8
作者：
Bolamba, D;Russ, KD;Durrant, BS
通讯作者：
Durrant, BS