Preimplantation Genetic Testing for Aneuploidy (PGT-A) Reveals High Levels of Chromosomal Errors in In Vivo-Derived Pig Embryos, with an Increased Incidence When Produced In Vitro.
Preimplantation Genetic Testing for Aneuploidy (PGT-A) Reveals High Levels of Chromosomal Errors in In Vivo-Derived Pig Embryos, with an Increased Incidence When Produced In Vitro.
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植入前非整倍体基因检测(PGT-A)在人类中广泛存在，但存在争议，并可改善牛的妊娠率和活产率。在猪身上，它为提高体外胚胎生产(IVP)提供了一种可能的解决方案，然而，染色体错误的发生率和来源仍未被探索。为了解决这一问题，我们使用了基于单核苷酸多态性的PGT-A算法，研究了101个活体来源的猪胚胎和体外受精的猪胚胎。体外受精与体外受精的囊胚率分别为79.7%和13.6%(P<；0.001)。在体外受精胚胎中，囊胚期的错误比卵裂期(4-细胞)的错误少(13.6%比40%，p=0.056)。还鉴定了1个雄核发育胚胎和2个孤雌生殖胚胎。三倍体在IVD胚胎中是最常见的错误(15.8%)，但仅在卵裂期观察到，而不是囊胚期，其次是全染色体非整倍体(9.9%)。IVP囊胚中孤雌生殖占32.8%，亚三倍体占25.0%，非整倍体占12.5%，单倍体占9.4%。每十头母猪中只有三头母猪产生了孤雌生殖囊胚，这表明可能存在供体效应。一般情况下，染色体异常的发生率很高，尤其是在IVP胚胎中，这表明了猪IVP成功率低的原因。所描述的方法提供了一种监测技术改进的手段，并建议未来应用PGT-A可能会提高胚胎移植的成功率。
Preimplantation genetic testing for aneuploidy (PGT-A) is widespread, but controversial, in humans and improves pregnancy and live birth rates in cattle. In pigs, it presents a possible solution to improve in vitro embryo production (IVP), however, the incidence and origin of chromosomal errors remains under-explored. To address this, we used single nucleotide polymorphism (SNP)-based PGT-A algorithms in 101 in vivo-derived (IVD) and 64 IVP porcine embryos. More errors were observed in IVP vs. IVD blastocysts (79.7% vs. 13.6% p < 0.001). In IVD embryos, fewer errors were found at blastocyst stage compared to cleavage (4-cell) stage (13.6% vs. 40%, p = 0.056). One androgenetic and two parthenogenetic embryos were also identified. Triploidy was the most common error in IVD embryos (15.8%), but only observed at cleavage, not blastocyst stage, followed by whole chromosome aneuploidy (9.9%). In IVP blastocysts, 32.8% were parthenogenetic, 25.0% (hypo-)triploid, 12.5% aneuploid, and 9.4% haploid. Parthenogenetic blastocysts arose from just three out of ten sows, suggesting a possible donor effect. The high incidence of chromosomal abnormalities in general, but in IVP embryos in particular, suggests an explanation for the low success of porcine IVP. The approaches described provide a means of monitoring technical improvements and suggest future application of PGT-A might improve embryo transfer success.
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