CReATe Fertility Centre, Toronto, Canada.
Department of Obstetrics and Gynecology, University of Toronto, Toronto, ON, Canada.
Sci Rep. 2019 Feb 27;9(1):2853. doi: 10.1038/s41598-019-39111-7.
Improved embryo prioritization is crucial in optimizing the results in assisted reproduction, especially in light of increasing utilization of elective single embryo transfers. Embryo prioritization is currently based on morphological criteria and in some cases incorporates preimplantation genetic testing for aneuploidy (PGT-A). Recent technological advances have enabled parallel genomic and transcriptomic assessment of a single cell. Adding transcriptomic analysis to PGT-A holds promise for better understanding early embryonic development and implantation, and for enhancing available embryo prioritization tools. Our aim was to develop a platform for parallel genomic and transcriptomic sequencing of a single trophectoderm (TE) biopsy, that could later be correlated with clinical outcomes. Twenty-five embryos donated for research were utilized; eight for initial development and optimization of our method, and seventeen to demonstrate clinical safety and reproducibility of this method. Our method achieved 100% concordance for ploidy status with that achieved by the classic PGT-A. All sequencing data exceeded quality control metrics. Transcriptomic sequencing data was sufficient for performing differential expression (DE) analysis. All biopsies expressed specific TE markers, further validating the accuracy of our method. Using PCA, samples clustered in euploid and aneuploid aggregates, highlighting the importance of controlling for ploidy in every transcriptomic assessment.
胚胎的优先选择在辅助生殖中优化结果至关重要，特别是在越来越多选择单胚胎移植的情况下。胚胎的优先选择目前基于形态学标准，在某些情况下还结合了非整倍体的胚胎植入前遗传学检测（PGT-A）。最近的技术进步使单细胞的基因组和转录组平行评估成为可能。将转录组分析添加到 PGT-A 中有望更好地了解早期胚胎发育和着床，并增强现有的胚胎优先选择工具。我们的目的是开发一种用于单个滋养外胚层（TE）活检的基因组和转录组平行测序的平台，该平台可与临床结果相关联。我们使用了 25 个用于研究的胚胎；其中 8 个用于初始开发和优化我们的方法，17 个用于证明这种方法的临床安全性和重现性。我们的方法与经典 PGT-A 获得的倍性状态的一致性达到 100%。所有测序数据都超过了质量控制指标。转录组测序数据足以进行差异表达（DE）分析。所有活检均表达特定的 TE 标志物，进一步验证了我们方法的准确性。使用 PCA，样品在整倍体和非整倍体聚集物中聚类，突出了在每个转录组评估中控制倍性的重要性。
