Effect of increased oxygen availability and astaxanthin supplementation on the growth, maturation and developmental competence of bovine oocytes derived from early antral follicles
Effect of increased oxygen availability and astaxanthin supplementation on the growth, maturation and developmental competence of bovine oocytes derived from early antral follicles
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DOI：
10.1016/j.theriogenology.2020.07.023
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发表时间：
2020-11-01
期刊：
影响因子：
2.8
通讯作者：
Nagano, Masashi
中科院分区：
文献类型：
--
作者：
Chelenga, Madalitso;Sakaguchi, Kenichiro;Nagano, Masashi
牛卵母细胞-卵丘-颗粒复合体（OCGC）的体外生长（IVG）培养通常使用常规的不透气培养装置进行12或14天。与体内卵泡发育相比，培养持续时间可能更长。在卵泡发育期间，卵泡从微血管接收氧气;然而，在使用常规不透气装置的培养下，氧气供应是有限的。本研究的目的是调查增加溶解氧的可用性，使用透气性（GP）培养装置与或不含抗氧化剂（虾青素，Ax）的补充8天的IVG培养系统的牛OCGCs来自早期窦卵泡的效果。我们在GP、补充有Ax的GP（GP + Ax）和常规不透气装置（对照）中培养OCGCs 8或12天。在所有培养条件下，培养8天的OCGC活力均显著高于培养12天的OCGC活力（p < 0.001），但组间差异不显著（p > 0.05）。在所有培养条件下，培养12天后OCGCs的胃窦形成率高于培养8天后的胃窦形成率（p < 0.001），并且无论是否补充Ax，对照组的胃窦形成率均显著高于GP组（p < 0.05）。第8天GP + Ax组、第8天对照组和第12天对照组的卵母细胞直径相似（p > 0.05）。体外培养8d的卵母细胞核成熟率，GP + Ax组显著高于对照组和GP组（p < 0.05），与培养12 d的卵母细胞核成熟率无显著差异（p > 0.05）。IVG培养第8天，GP + Ax组OCGCs活性氧产生量明显低于GP组和对照组（p < 0.05）。IVG卵母细胞培养8天后发育成囊胚，所有处理组的卵裂率和囊胚率相似。然而，在所有组中，体内生长的卵母细胞的卵裂率和囊胚率显著高于IVG卵母细胞（p < 0.05）。本研究表明，增加氧气的可用性，使用GP培养装置与Ax补充促进卵母细胞的生长和成熟能力，但抑制颗粒细胞的增殖和胃窦的形成相比，传统的不透气的培养装置，和OCGCs可以达到8天的IVG培养后的发育能力。(C)2020爱思唯尔公司All rights reserved.
In vitro growth (IVG) culture of bovine oocyte-cumulus-granulosa complexes (OCGCs) is generally carried out for 12 or 14 days using conventional gas impermeable culture devices. The culture duration may be longer compared to follicular development in vivo. During follicular development, follicles receive oxygen from micro vessels; however, oxygen supply is limited under the culture using conventional gas impermeable devices. The purpose of this study was to investigate the effect of increasing dissolved oxygen availability using a gas permeable (GP) culture device with or without antioxidant (astaxanthin, Ax) supplementation on 8-day IVG culture systems for bovine OCGCs derived from early antral follicles. We cultured OCGCs in GP, GP supplemented with Ax (GP + Ax), and a conventional gas impermeable device (control) for 8 or 12 days. OCGC viability were significantly higher when cultured for 8 days than 12 days (p < 0.001) in all culture condition, but significant difference was not observed between groups (p > 0.05). Antrum formation rates of OCGCs were higher after 12 days than 8 days of culture in all culture condition (p < 0.001) and were significantly higher in the control than GP groups regardless of Ax supplementation (p < 0.05). Oocyte diameters were similar among day-8 GP + Ax, day-8 control and day-12 control groups (p > 0.05). Nuclear maturation rates of oocytes grown in vitro for 8 days were significantly higher in the GP + Ax group than in the control and the GP groups (p < 0.05) and similar to oocytes grown for 12 days regardless of the culture conditions (p > 0.05). The generation of reactive oxygen species in OCGCs on day 8 of IVG culture was significantly lower in the GP + Ax group than those of the GP and control groups (p < 0.05). IVG oocytes after eight days of culture developed into blastocysts, and the cleavage and blastocyst rates were similar in all treatment groups. However, in vivo-grown oocytes had significantly higher (p < 0.05) cleavage and blastocyst rates than the IVG oocytes in all groups. The present study demonstrates that increased oxygen availability using a GP culture device with Ax supplementation promotes oocyte growth and maturation competence but inhibits proliferation of granulosa cells and antrum formation compared with a conventional gas impermeable culture device, and that OCGCs can attain developmental competence after 8 days of IVG culture. (C) 2020 Elsevier Inc. All rights reserved.