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Effects of maturational age of recipient oocytes and activation conditions on the development of porcine fetal fibroblast nuclear transfer embryos

机译:受体卵母细胞成熟年龄和激活条件对猪胎儿成纤维细胞核移植胚胎发育的影响

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摘要

This study was conducted to evaluate the nuclear remodeling patterns and the developmental potential of porcine fetal fibroblast nuclear transfer embryos (NTs) following the maturational age of recipient oocytes and activation conditions. Donor cells were transferred into the enucleated oocytes that were matured for 36 or 44h. Electrofused embryos were cultured in PZM-3 for 6 days without activation treatment (EF group). Some of these embryos were additionally activated by electric stimulus (ES; EF+ES group) or a combination of ES and DMAP (EF+ES+D group) before culture. The reconstituted embryos were fixed 2.5h after fusion to evaluate the nuclear remodeling patterns. The nuclear remodeling pattern of NTs reconstituted with 44 h-matured recipients showed a tendency to form a pronucleus-like structure, while that of NTs reconstituted with 36 h-matured recipients showed a tendency to undergo a premature chromosome condensation (PCC) and form one set of chromatin clump. In EF+ES+D group, blastocyst development was significantly increased regardless of maturational age of recipient oocytes (P<0.05). The result indicates that additional activation treatment is necessary to induce the activation of embryos reconstituted with 36 h-matured recipients, and treatment with the combination of electrical stimuli and DMAP could enhance the blastocyst formation rate of porcine NTs reconstituted with both 36 h- and 44 h-matured recipient oocytes.
机译:这项研究的目的是评估受体卵母细胞的成熟年龄和激活条件后,猪胎儿成纤维细胞核移植胚胎(NTs)的核重塑模式和发展潜力。将供体细胞转移到去核的卵母细胞中,该卵母细胞成熟36或44h。在未经激活处理的情况下,将电融合的胚胎在PZM-3中培养6天(EF组)。在培养之前,通过电刺激(ES; EF + ES组)或ES和DMAP的组合(EF + ES + D组)额外激活其中一些胚胎。融合后2.5小时将重构的胚固定,以评估核重构模式。用44h成熟的受体重组的NTs的核重塑模式显示出形成核样结构的趋势,而用36h成熟的受体重组的NTs的核重塑模式显示出发生早染色体冷凝(PCC)并形成一个核样结构的趋势。组的染色质团块。在EF + ES + D组,无论受体卵母细胞的成熟年龄如何,胚泡发育均显着增加(P <0.05)。结果表明,需要额外的活化处理来诱导36 h成熟受体重构的胚胎的活化,电刺激和DMAP的组合处理可以提高36 h和44重构的猪NTs的胚泡形成率h成熟的受体卵母细胞。

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