Effect of the addition of insulin-transferrin-selenium and/or L-ascorbic acid to the in vitro maturation of prepubertal bovine oocytes on cytoplasmic maturation and embryo development
Effect of the addition of insulin-transferrin-selenium and/or L-ascorbic acid to the in vitro maturation of prepubertal bovine oocytes on cytoplasmic maturation and embryo development
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DOI：
10.1016/j.theriogenology.2010.06.003
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发表时间：
2010-11-01
期刊：
影响因子：
2.8
通讯作者：
Mogas, T.
中科院分区：
文献类型：
--
作者：
Cordova, B.;Morato, R.;Mogas, T.
本研究考察了在常规培养基中添加胰岛素-转铁蛋白-硒（ITS）和/或l -抗坏血酸（ASC）对成熟的青春期前小牛卵母细胞染色体组织、皮质颗粒（CG）分布和胚胎发育到囊胚期的影响。积云卵母细胞复合物（COCs）在含有PVA和EGF（对照）的TCM 199培养基中成熟，并在38.5℃、5% CO2气氛中添加ITS和/或ASC，培养12或24 h。在ITS + ASC或ASC作用下成熟12 h的小牛卵母细胞外周分布的CG百分比（分别为83.3%和86.2%）显著高于对照卵母细胞（71.4%）或单独使用ITS成熟的卵母细胞（71.4%）。未检测到对染色体组织的影响。相反，添加24 h不影响CG分布模式，而添加ASC使卵母细胞染色体正常排列的百分比（72.9%）显著高于对照组（58.7%）。在48 hpi时，无论处理时间如何，都观察到相似的裂解率。然而，ITS + ASC存在12 h后，囊胚率明显高于其他组。添加ITS或ITS + ASC 24 h对获得囊胚的比例没有显著影响，而ASC的存在显著降低了胚胎发育到囊胚期的比例。我们的数据表明，在体外成熟的前12小时补充ITS和l -抗坏血酸可以促进细胞质成熟和由青春期前小牛卵母细胞产生的胚胎的发育能力。(C) 2010爱思唯尔公司版权所有。
This study examines the effects of adding insulin-transferrin-selenium (ITS) and/or L-ascorbic acid (ASC) to a conventional medium for maturing prepubertal calf oocytes on chromosome organization, cortical granule (CG) distribution, and embryo development to the blastocyst stage. Cumulus-oocyte complexes (COCs) were matured in medium TCM 199 containing PVA and EGF (control), and supplemented with ITS and/or ASC for 12 or 24 h at 38.5 degrees C in a 5% CO2 atmosphere. Calf oocytes matured with ITS + ASC or ASC for 12 h showed significantly higher percentages of peripherally distributed CG (83.3% and 86.2% respectively) than control oocytes (71.4%) or those matured with ITS alone (71.4%). No effects on chromosome organization were detected. Conversely, 24 h of supplementation did not affect CG distribution patterns, while the addition of ASC gave rise to significantly higher percentages of oocytes showing a normal alignment of their chromosomes (72.9%) compared to controls (58.7%). At 48 hpi, similar cleavage rates were observed among treatments regardless of the treatment time. However, the presence of ITS + ASC for 12 h rendered significantly higher blastocyst rates than those recorded in the remaining groups. Supplementation for 24 h with ITS or ITS + ASC had no significant effects on the percentage of blastocysts obtained, while the presence of ASC significantly reduced the proportions of embryos developing to the blastocyst stage. Our data suggest that ITS plus L-ascorbic acid supplementation during the first 12 h of in vitro maturation improves cytoplasm maturation and the developmental competence of embryos produced from prepubertal calf oocytes. (C) 2010 Elsevier Inc. All rights reserved.