Optimized Protocols for In Vitro Maturation of Rat Oocytes Dramatically Improve Their Developmental Competence to a Level Similar to That of Ovulated Oocytes
Optimized Protocols for In Vitro Maturation of Rat Oocytes Dramatically Improve Their Developmental Competence to a Level Similar to That of Ovulated Oocytes
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DOI：
10.1089/cell.2015.0055
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发表时间：
2016-02-01
影响因子：
1.6
通讯作者：
Tan, Jing-He
中科院分区：
文献类型：
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作者：
Jiao, Guang-Zhong;Cui, Wei;Tan, Jing-He
体外成熟（IVM）卵母细胞的发育能力明显低于其体内成熟（IVO）对应物，这表明需要优化不同物种的IVM方案。关于大鼠卵母细胞体外成熟的研究很少，与其他物种的报道相比，提高卵浆成熟的尝试更少。此外，众所周知，大鼠卵母细胞在离开输卵管后进行自发激活（SA）;然而，IVM大鼠卵母细胞是否具有比IVO卵母细胞更低的SA率，并且可以潜在地用于核移植尚不清楚。在本研究中，我们研究了成熟程序对体外成熟大鼠卵母细胞胞质成熟的影响，并观察了使用体外成熟大鼠卵母细胞降低SA的可能性。使用多种标记物评估卵质成熟，包括植入前和植入后发育、减数分裂进程、CG再分布、氧化还原状态以及发育潜力和发育障碍相关基因的表达。结果表明，添加半胱胺/胱氨酸的TCM-199培养液和卵丘细胞单层可显著提高大鼠卵母细胞的发育能力，并使着床前和着床后的发育及其他卵质成熟指标达到与排卵卵母细胞相似的水平。在核成熟后的相同时间间隔内，IVM卵母细胞的SA率显著低于IVO卵母细胞。总之，我们优化了大鼠卵母细胞体外成熟的方案，使卵浆成熟到与排卵卵母细胞相似的水平。结果表明，体外成熟大鼠卵母细胞可用于降低SA大鼠克隆。
The developmental capacity of in vitro-matured (IVM) oocytes is markedly lower than that of their in vivo-matured (IVO) counterparts, suggesting the need for optimization of IVM protocols in different species. There are few studies on IVM of rat oocytes, and there are even fewer attempts to improve ooplasmic maturation compared to those reported in other species. Furthermore, rat oocytes are well known to undergo spontaneous activation (SA) after leaving the oviduct; however, whether IVM rat oocytes have lower SA rates than IVO oocytes and can potentially be used for nuclear transfer is unknown. In this study, we investigated the effects of maturation protocols on cytoplasmic maturation of IVM rat oocytes and observed the possibility to reduce SA by using IVM rat oocytes. Ooplasmic maturation was assessed using multiple markers, including pre- and postimplantation development, meiotic progression, CG redistribution, redox state, and the expression of developmental potential-and apoptosis-related genes. The results showed that the best protocol consisting of modified Tissue Culture Medium-199 (TCM-199) supplemented with cysteamine/cystine and the cumulus cell monolayer dramatically improved the developmental competence of rat oocytes and supported both pre-and postimplantation development and other ooplasmic maturation makers to levels similar to that observed in ovulated oocytes. Rates of SA were significantly lower in IVM oocytes than in IVO oocytes when observed at the same intervals after nuclear maturation. In conclusion, we have optimized protocols for IVM of rat oocytes that sustain ooplasmic maturation to a level similar to ovulated oocytes. The results suggest that IVM rat oocytes might be used to reduce SA for rat cloning.