关键词: Humans In vitro maturation Nuclear transfer Oocytes Polar bodies Spindle apparatus

来  源:   DOI:10.5653/cerm.2023.05939   PDF(Pubmed)

Abstract:
OBJECTIVE: Reconstructed oocytes after polar body genome transfer constitute a potential therapeutic option for patients with a history of embryo fragmentation and advanced maternal age. However, the rescue of genetic material from the first polar body (PB1) through introduction into the donor cytoplasm is not yet ready for clinical application.
METHODS: Eighty-five oocytes were obtained following in vitro maturation (IVM) and divided into two groups: PB1 nuclear transfer (PB1NT; n=54) and control (n=31). Following enucleation and PB1 genomic transfer, PB1 fusion was assessed. Subsequently, all fused oocytes underwent intracytoplasmic sperm injection (ICSI) and were cultured in an incubator under a time-lapse monitoring system to evaluate fertilization, embryonic morphokinetic parameters, and cleavage patterns.
RESULTS: Following enucleation and fusion, 77.14% of oocytes survived, and 92.59% of polar bodies (PBs) fused. However, the normal fertilization rate was lower in the PB1NT group than in the control group (56.41% vs. 92%, p=0.002). No significant differences were observed in embryo kinetics between the groups, but a significant difference was detected in embryo developmental arrest after the four-cell stage, along with abnormal cleavage division in the PB1NT group. This was followed by significant between-group differences in the implantation potential rate and euploidy status. Most embryos in the PB1NT group had at least one abnormal cleavage division (93.3%, p=0.001).
CONCLUSIONS: Fresh PB1NT oocytes successfully produced normal zygotes following PB fusion and ICSI in IVM oocytes. However, this was accompanied by low efficiency in developing into cleavage embryos, along with an increase in abnormal cleavage patterns.
摘要:
目的:极体基因组转移后的卵母细胞重建是有胚胎碎片史和高龄患者的潜在治疗选择。然而,通过引入供体细胞质从第一极体(PB1)中拯救遗传物质尚未准备好临床应用。
方法:体外成熟(IVM)后获得85个卵母细胞,分为两组:PB1核移植(PB1NT;n=54)和对照组(n=31)。摘除和PB1基因组转移后,评估PB1融合。随后,所有融合的卵母细胞均接受胞浆内单精子注射(ICSI),并在延时监测系统下在培养箱中培养以评估受精,胚胎形态动力学参数,和乳沟模式。
结果:摘除和融合后,77.14%的卵母细胞存活,和92.59%的极体(PBs)融合。然而,PB1NT组的正常受精率低于对照组(56.41%vs.92%,p=0.002)。两组之间的胚胎动力学没有显着差异,但是在四细胞阶段后的胚胎发育停滞中发现了显着差异,伴随着PB1NT组的异常卵裂分裂。随后是植入电位率和整倍体状态的显着组间差异。PB1NT组中的大多数胚胎至少有一次异常卵裂分裂(93.3%,p=0.001)。
结论:新鲜的PB1NT卵母细胞在IVM卵母细胞中成功地在PB融合和ICSI后产生正常受精卵。然而,这伴随着发育成卵裂胚胎的效率低下,随着异常卵裂模式的增加。
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