Greco Ermanno, Biricik Anil, Cotarelo Rocio P, Iammarone Elisabetta, Rubino Patrizia, Tesarik Jan, Fiorentino Francesco, Minasi Maria Giulia
Centre for Reproductive Medicine, European Hospital, Rome, Italy.
Genoma, Molecular Genetics Laboratory, Rome, Italy.
Springerplus. 2015 Jan 14;4:22. doi: 10.1186/s40064-015-0788-y. eCollection 2015.
Preimplantation genetic diagnosis and/or screening (PGD/PGS) allow the assessment of the genetic health of an embryo before transferring it into the uterus. These techniques require the removal of cellular material (polar bodies, blastomere(s) or trophectoderm cells) in order to perform the proper genetic analysis. We report the implantation and live birth outcome of a vitrified-warmed blastocyst developed after triple biopsy and double vitrification procedures at oocyte, cleavage embryo and blastocyst stage.
An infertile couple, with family history of β-thalassemia, searched for IVF procedure and PGD. First polar bodies biopsy with subsequent vitrification was uninformative due to meiotic crossing-over, so oocytes were inseminated after warming. Two embryos were obtained and blastomere biopsy was performed on day 3 with inconclusive results on their genetic status. Their culture resulted in one expanded blastocyst stage on day 7 that underwent trophectoderm biopsy and vitrification. This embryo showed to be normal. It was then warmed and transferred in an artificial cycle.
Preconception genetic analysis by removal and analysis of the first polar body is technically possible, but the genetic information that we can obtain at this stage may be limited and the oocytes to be inseminated is not predictable. Compared to blastomere biopsy, trophectoderm biopsy has more diagnostic efficiency with respect to both chromosomal mosaicism and PCR accuracy, reducing the problems of amplification failure and allele drop out. Moreover, embryos biopsied at the cleavage stage seem to have lower implantation rate than biopsied blastocyst.
This is the first case report of a live birth obtained from a three step biopsy and double vitrification procedures of a blastocyst. This case report seems also to suggest the harmlessness of all these procedures if carefully performed by a skilled biologist in an IVF lab with quality management system. Finally, our study highlight that blastocyst cryopreserved on day 7 have clinically important potential and embryos that not reach blastocyst stage on day 6 should not to be discharged because they may result in an ongoing pregnancy.
胚胎植入前基因诊断和/或筛查(PGD/PGS)可在胚胎植入子宫前评估其基因健康状况。这些技术需要去除细胞物质(极体、卵裂球或滋养外胚层细胞)以进行适当的基因分析。我们报告了在卵母细胞、卵裂期胚胎和囊胚阶段经过三次活检和两次玻璃化程序后发育而成的玻璃化-复温囊胚的植入和活产结局。
一对有β地中海贫血家族史的不孕夫妇寻求体外受精(IVF)和PGD。由于减数分裂交叉互换,首次极体活检及随后的玻璃化处理未提供有用信息,因此在复温后对卵母细胞进行了授精。获得了两个胚胎,并在第3天进行了卵裂球活检,但关于其基因状态的结果不明确。它们培养至第7天形成了一个扩张期囊胚,该囊胚接受了滋养外胚层活检和玻璃化处理。这个胚胎显示正常。然后将其复温并在人工周期中进行移植。
通过去除并分析第一极体进行孕前基因分析在技术上是可行的,但我们在这个阶段能够获得的基因信息可能有限,并且有待授精的卵母细胞不可预测。与卵裂球活检相比,滋养外胚层活检在染色体嵌合和聚合酶链反应(PCR)准确性方面具有更高的诊断效率,减少了扩增失败和等位基因脱失的问题。此外,在卵裂期进行活检的胚胎似乎比活检囊胚的植入率更低。
这是首例关于通过对囊胚进行三步活检和两次玻璃化程序获得活产的病例报告。该病例报告似乎还表明,如果由熟练的生物学家在具有质量管理体系的IVF实验室中仔细操作所有这些程序,它们是无害的。最后,我们的研究强调,第7天冷冻保存的囊胚具有重要的临床潜力,并且在第6天未达到囊胚阶段的胚胎不应被放弃,因为它们可能会导致持续妊娠。