Zorina I M, Eldarov C M, Yarigina S A, Makarova N P, Trofimov D Yu, Smolnikova V Yu, Kalinina E A, Bobrov M Yu
Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Moscow, Russia.
Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Moscow, Russia; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia.
Biomed Khim. 2017 Oct;63(5):385-391. doi: 10.18097/PBMC20176305385.
The aim of this study was to determine the changes of metabolomic profiles in embryonic culture media (ECM) for the evaluation of quality and implantation potential of human embryos. ECM (n=163) were collected on day 5 before transfer or cryopreservation. Some embryos were used in preimplantation genetic screening for detection of aneuploidy karyotypes. Samples were subdivided into groups according to embryo morphological classification (by Gardner), genetic analysis and implantation data. ECM were extracted with methanol, precipitates were separated by centrifugation and metabolite production of individual embryo was analysed by LC-MS (the positive ion mode). After peak detection and retention time alignment, data were analysed using the PCA algorithm. MS fingerprinting analysis of embryo culture medium showed significant differences between morphologically divided groups. Intragroup comparisons did not reveal differences between subclasses. Genetic screening of embryos revealed 33 aneuploid karyotypes. It was shown that chromosome number did not affect the metabolite profiles comparing with the normal group. The culture media of embryos that were positive or negative for successful implantation showed specific signatures that allowed to distinguish embryos with different outcomes.The characterization of ECMs by LC-MS may facilitate more accurate selection of the best embryo for the implantation, improving single-embryo transfer and thus eliminating the risk and undesirable effects of multiple pregnancies.
本研究的目的是确定胚胎培养基（ECM）中代谢组学谱的变化，以评估人类胚胎的质量和着床潜力。在移植或冷冻保存前的第5天收集ECM（n = 163）。一些胚胎用于植入前基因筛查以检测非整倍体核型。根据胚胎形态学分类（由Gardner法）、基因分析和着床数据将样本分组。用甲醇提取ECM，通过离心分离沉淀物，并通过液相色谱 - 质谱联用仪（正离子模式）分析单个胚胎的代谢产物生成情况。在进行峰检测和保留时间校准后，使用主成分分析（PCA）算法分析数据。胚胎培养基的质谱指纹图谱分析显示，形态学分组之间存在显著差异。组内比较未发现亚类之间存在差异。胚胎的基因筛查发现了33种非整倍体核型。结果表明，与正常组相比，染色体数目不影响代谢产物谱。着床成功或失败的胚胎培养基显示出特定特征，能够区分具有不同结局的胚胎。通过液相色谱 - 质谱联用仪对ECM进行表征，可能有助于更准确地选择最佳着床胚胎，改善单胚胎移植，从而消除多胎妊娠的风险和不良影响。
