Hanging Drop Monoculture for Selection of Optimal Antioxidants During In Vitro Maturation of Porcine Oocytes
Hanging Drop Monoculture for Selection of Optimal Antioxidants During In Vitro Maturation of Porcine Oocytes
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DOI：
10.1111/rda.12289
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发表时间：
2014-04-01
影响因子：
1.7
通讯作者：
Tanemura, K.
中科院分区：
文献类型：
--
作者：
Ishikawa, S.;Machida, R.;Tanemura, K.
研究了三种不同作用机制的抗氧化剂对猪卵母细胞体外成熟的影响。单卵母细胞单培养使用悬滴（HD）系统具有一些优点，如提高分析效率带来的样品数量比一滴培养的卵母细胞的数量少。配体对单个卵母细胞的直接作用也可以在不考虑其他卵母细胞的旁分泌因子的情况下检测到。在预培养22 h后，将裸露的卵母细胞与0.01和0.1g/ml的L-肉毒碱（LC）、乳铁蛋白（LF）或萝卜硫素（SF）在存在/不存在由补充H2 O2诱导的氧化应激的情况下培养22 h，以评估抗氧化应激对核成熟的降低作用。结果表明，与LC和SF相比，LF在较低浓度（0.01g/ml）下就能有效降低猪卵母细胞体外成熟过程中的氧化应激，表明LF是一种更有效的抗氧化剂。此外，LF也增加成熟率，即使在没有过氧化氢的培养。我们的研究结果清楚地表明，HD单培养系统是有用的筛选影响猪卵母细胞培养的物质。
Contents We analysed the effect of three antioxidants that have different functional mechanisms on the in vitro maturation (IVM) of porcine oocytes. Single oocyte monoculture using the hanging drop (HD) system has some advantages such as improving analysis efficiency brought by the smaller number of samples than the number of oocytes cultured in one drop. Direct effects of ligands on single oocytes could also be detected without considering the effects of paracrine factors from other oocytes. After 22h of pre-culture, denuded oocytes were cultured for 22h with 0.01 and 0.1g/ml of L-carnitine (LC), lactoferrin (LF) or sulforaphane (SF) in the presence/non-presence of oxidant stress induced by H2O2 supplementation to evaluate the reducing effects against oxidative stress on nuclear maturation. As a result, compared with LC and SF, LF showed effective reduction in oxidative stress at a lower concentration (0.01g/ml), suggesting that LF is a more effective antioxidant in porcine oocyte IVM. Additionally, LF also increased maturation rate even in culture without H2O2. Our results clearly suggest that the HD monoculture system is useful for screening the substances that affect porcine oocyte culture.