The developmental potential of parthenogenetic and somatic cell nuclear-transferred rat oocytes in vitro.

Shigetoshi Mizumoto, Yoko Kato, Yukio Tsunoda
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引用次数: 15

Abstract

We examined the optimal conditions for somatic cell nuclear transfer (SCNT) in rat oocytes. First, we compared the effects of two types of inhibitors of spontaneous activation, MG132 and demecolcine, on the developmental potential of parthenogenetic oocytes. The potential of activated oocytes to develop into blastocysts significantly decreased 2 h after oocyte recovery (77% vs. 7%). The developmental potential of oocytes preserved in MG132-supplemented medium for 1 to 4 h was high (62% to 77%), but the potential of those preserved in demecolcine-supplemented medium for 3 and 4 h was low (77% vs. 41% and 37%, respectively). Second, the effect of the duration of parthenogenetic activation on the developmental potential was examined. When oocytes preserved in MG132 for 4 h were treated with 10 mM strontium for 5 or 6 h, the potential of activated oocytes to develop into blastocysts was high (78% and 70%, respectively). Using the optimal conditions for parthenogenetic activation, we examined the potential of rat enucleated oocytes receiving cumulus cells to develop into blastocysts. In contrast to parthenogenotes, the potential of SCNT rat oocytes to develop into blastocysts was low (2%) even if then oocytes were treated with the histone deacetylation inhibitor trichostatin A. The reason for the low developmental potential of rat SCNT oocytes is discussed.

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孤雌和体细胞核移植大鼠卵母细胞的体外发育潜力。
我们研究了大鼠卵母细胞体细胞核移植(SCNT)的最佳条件。首先,我们比较了两种自发活化抑制剂MG132和去美可林对孤雌卵母细胞发育潜能的影响。卵母细胞恢复后2小时,活化卵母细胞发育成囊胚的潜力显著降低(77% vs. 7%)。在mg132培养基中保存1 ~ 4小时的卵母细胞发育潜力较高(62% ~ 77%),而在deecolcine培养基中保存3 ~ 4小时的卵母细胞发育潜力较低(77%,分别为41%和37%)。其次,研究了孤雌生殖激活时间对发育潜能的影响。在MG132中保存4 h的卵母细胞用10 mM锶处理5或6 h后,活化卵母细胞发育成囊胚的可能性很高(分别为78%和70%)。利用最佳的孤雌激活条件,研究了接受卵丘细胞的大鼠去核卵母细胞发育成囊胚的潜力。与孤雌生殖相比,SCNT大鼠卵母细胞发育成囊胚的潜力很低(2%),即使用组蛋白去乙酰化抑制剂曲古抑素a处理。本文讨论了SCNT大鼠卵母细胞发育潜力低的原因。
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