Successful implantation of in vitro-matured, electro-activated oocytes in the pig
Successful implantation of in vitro-matured, electro-activated oocytes in the pig
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DOI：
10.1016/s0093-691x(00)00256-9
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发表时间：
2000-03-15
期刊：
影响因子：
2.8
通讯作者：
Kato, S
中科院分区：
文献类型：
--
作者：
Kure-Bayashi, S;Miyake, M;Kato, S
在本研究中，我们从体外成熟的卵母细胞经过简单的激活过程获得了孤雌生殖的猪胎儿，以评估其在体内的发育限制。从当地屠宰场收集的母猪卵泡卵母细胞成熟48 h。将它们以1，500 V/cm的直流电进行100 μ sec的单一方形脉冲，然后用5 μ g/mL细胞松弛素B处理4小时，以获得活化的二倍体卵母细胞。在改良的Whitten培养基中培养二倍体直至转移。将分裂至2-和3-至4-细胞期的二倍体转移至受体的输卵管中。在激活后17、18、19、24和29 d，8项试验中有6项回收了活的和/或死的孤雌生殖胎仔。移植的胎儿占二倍体的总比例为31.3%（62/198）。当激活后19 d回收胎仔时，发育成胎仔的比例为71%（15/21）。然而，我们的结果表明，妊娠期超过19天导致这些比例下降到45%（18/40），在24天和18%（7/40），在29天。在激活后19、24和29 d，几乎所有的胎儿心脏都在跳动。因此，孤雌生殖的猪二倍体发育到至少超过早期心脏跳动阶段的肢芽形成阶段。在一些胎仔上也可从外部观察到流产。在某些病例中观察到心脏和肝脏中形成囊肿样结构，头部和头状脑发育不足。(C)2000年，Elsevier Science Inc.
In the present study, we derived parthenogenetic porcine fetuses from in vitro-matured oocytes following a simple activation process in order to evaluate their developmental limitations in-vivo. Follicular oocytes collected from gilts at local slaughterhouses were matured for 48 h. They were subjected to a single square pulse of direct current for 100 mu sec at 1,500 V/cm and then treated with 5 mu g/mL cytochalasin B for 4 h to obtain activated diploid oocytes. The diploids were cultured in modified Whitten's medium until transfer. Diploids which had cleaved to the 2- and 3- to 4-cell stages were transferred to oviducts of recipients. Live and/or dead parthenogenetic fetuses were recovered in 6 of 8 trials at 17, 18, 19, 24 and 29 d post activation. The total proportion of fetuses to transferred diploids was 31.3% (62/198). When fetuses were recovered at 19 d post activation, the proportion of development into fetuses was 71% (15/21). Our results, however, suggest that periods of gestation longer than 19 d resulted in a decrease of these proportions to 45% (18/40) at 24 d and to 18% (7/40) at 29 d. The hearts were beating in nearly all of the fetuses recovered at 19, 24 and 29 d post activation. Thus, parthenogenetic porcine diploids developed to at least the stage of limb-bud formation beyond the early heart-beating stage. Abnormalities were also externally visible on some fetuses. Formation of cyst-like structures in the heart and liver, and insufficient development of the head region and acephali were observed in some cases. (C) 2000 by Elsevier Science Inc.