目的:我们的目的是确定极体基因组转移(PBT)后人类重建卵母细胞的发育潜力,并报道一例具有多个周期的严重胚胎碎片的女性。
方法:将新鲜和冷冻保存的第一极体(PB1s)转移到去核中期II卵母细胞(PB1T)中,而从受精的卵母细胞中取出新鲜的PB2s,并代替供体受精卵中的雌性原核使用。重建的卵母细胞接受胞浆内精子注射(ICSI)并培养至胚泡。通过下一代测序(NGS)筛选PBT衍生的胚泡的生物滋养外胚层细胞的染色体。然后,冷冻保存的PB1T在一名有几个周期广泛胚胎破碎史的女性中进行,并且对来自PB1T的囊胚进行非整倍性筛查,但未转移给患者。
结果:新鲜和冷冻保存的PB1T与对照卵母细胞的正常受精率和胚泡形成率没有显着差异。在三个新鲜和三个冷冻保存的PB1T来源的胚泡中,两个和一个胚泡分别表现出正常的二倍体。相比之下,17个PB2转移产生了16个两个原核(2PN)受精卵,每个都有一个正常和一个小尺寸的原核,没有胚泡形成。在女病人身上,在第四个周期中,通过ICSI授精18个卵母细胞,并对PB1s进行活检。尽管从患者自己的卵母细胞发育的胚胎显示出严重的碎片,PB1T后重建的卵母细胞产生了三个染色体正常的胚泡。
结论:正常的胚泡可以在PB1T后从人的重建卵母细胞发育。首次PB转移的应用可能对胚胎发育不良和过度碎片化病史的患者有益。
OBJECTIVE: We aimed to determine the developmental potential of human reconstructed oocytes after polar body genome transfer (PBT) and to report the
case of a woman with multiple cycles of severe embryo fragmentation.
METHODS: Fresh and cryopreserved first polar bodies (PB1s) were transferred to enucleated metaphase II oocytes (PB1T), while fresh PB2s were removed from fertilized oocytes and used instead of the female pronucleus in donor zygotes. Reconstructed oocytes underwent intracytoplasmic sperm injection (ICSI) and were cultured to blastocyst. Biopsied trophectoderm cells of PBT-derived blastocysts were screened for chromosomes by next-generation sequencing (NGS). Then, cryopreserved PB1T was carried out in one woman with a history of several cycles of extensive embryo fragmentation, and the blastocysts derived from PB1T were screened for aneuploidy but not transferred to the patient.
RESULTS: There were no significant differences in the rates of normal fertilization and blastocyst formation between fresh and cryopreserved PB1T and control oocytes. Of the three fresh and three cryopreserved PB1T-derived blastocysts, two and one blastocysts exhibited normal diploidy respectively. In contrast, 17 PB2 transfers yielded 16 two pronuclei (2PN) zygotes with one normal and one small-sized pronucleus each and no blastocyst formation. In the female patient, 18 oocytes were inseminated by ICSI in the fourth cycle and the PB1s were biopsied. Although the embryos developed from the patient\'s own oocytes showed severe fragmentation, the oocytes reconstructed after PB1T produced three chromosomally normal blastocysts.
CONCLUSIONS: Normal blastocysts can develop from human reconstructed oocytes after PB1T. The application of the first PB transfers may be beneficial to patients with a history of poor embryo development and excessive fragmentation.