LC-MS Analysis Revealed the Significantly Different Metabolic Profiles in Spent Culture Media of Human Embryos with Distinct Morphology, Karyotype and Implantation Outcomes.

Elena Kalinina,Svetlana Yarigina,Luiza Ibragimova,Gennady Sukhikh,Mikhail Bobrov,Veronika Smolnikova,Nataliya Makarova,Vitaliy Chagovets,Chupalav Eldarov,Alina Gamisonia,Victor Zgoda

doi:10.3390/ijms23052706

Abstract

In this study we evaluated possible differences in metabolomic profiles of spent embryo culture media (SECM) of human embryos with distinct morphology, karyotype, and implantation outcomes. A total of 153 samples from embryos of patients undergoing in vitro fertilization (IVF) programs were collected and analyzed by HPLC-MS. Metabolomic profiling and statistical analysis revealed clear clustering of day five SECM from embryos with different morphological classes and karyotype. Profiling of day five SECM from embryos with different implantation outcomes showed 241 significantly changed molecular ions in SECM of successfully implanted embryos. Separate analysis of paired SECM samples on days three and five revealed 46 and 29 molecular signatures respectively, significantly differing in culture media of embryos with a successful outcome. Pathway enrichment analysis suggests certain amino acids, vitamins, and lipid metabolic pathways to be crucial for embryo implantation. Differences between embryos with distinct implantation potential are detectable on the third and fifth day of cultivation that may allow the application of culture medium analysis in different transfer protocols for both fresh and cryopreserved embryos. A combination of traditional morphological criteria with metabolic profiling of SECM may increase implantation rates in assisted reproductive technology programs as well as improve our knowledge of the human embryo metabolism in the early stages of development.

Highlights

IntroductionWhile the number of assisted reproductive technologies (ART) cycles steadily increase worldwide, live birth rates vary significantly over different regions—from 12–51% for fresh embryos to 18–57% for cryopreserved ones [2]
The elective single-embryo transfer has been shown in several studies to be the recommended method compared to multiple embryo transfer, since it demonstrates a similar birth rate and is devoid of disadvantages associated with multiple pregnancies [41–43]
This method requires a comprehensive assessment of the viability and implantation potential of a single embryo, as there is no room for error

Summary

Introduction

While the number of ART cycles steadily increase worldwide, live birth rates vary significantly over different regions—from 12–51% for fresh embryos to 18–57% for cryopreserved ones [2]. One of the ways to increase ART efficiency is double embryo transfer [3–6], but this approach has well-known risks of multiple pregnancies, associated with different complications for infant and mother health [7,8], which have tendency to increase with age [9,10]. Over the past two decades, there has been a shift towards the elective single embryo transfer (eSET) protocols [11]. While this approach mitigates risks associated with multi-pregnancy, it requires developing an accurate strategy of embryo quality assessment, selection, and transfer. The question was raised of when the embryo should be transferred

Methods

Results

Conclusion