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Noninvasive assessment of embryo quality in human in vitro fertilization : metabolomic profiling of embryo culture media with Raman spectroscopy Asghari Roodsari, Alaleh


Introduction: Light microscopy has remained the primary tool for the assessment of embryo quality and the selection of embryos for transfer in clinical IVF practice. Recent studies have suggested that metabolomic profiling of embryo culture media can distinguish human embryos with better implantation potential. We therefore undertook the following study to further assess the usefulness of metabolomic profiling “spent” embryo culture medium using Raman spectroscopy. Methods: Patients undergoing IVF+/-ICSI treatment from the UBC Centre for Reproductive Health were recruited for study. Demographic and clinical information was collected. As part of routine clinical procedures, embryos were individually cultured in G1 media from Day 1-3 and in G2 media from Day 4-6. G1 and G2 culture medium (vitrolife, Englewood, CO) introduced specifically for cleavage embryo and blastocyst culture respectively. Embryo-free G1 and G2 droplets were placed alongside the embryo-containing ones as controls. For the study, fresh droplets of spent and control culture media were individually collected on Day 3 and 6 and prepared for assessment by Raman spectroscopy. The assessment score under light microscopy of the corresponding embryo and its fate were recorded for comparison and correlation. To validate the detection limits of Raman spectroscopy a wide range of glucose and glycine concentrations between 0 and 500 mM in distilled water were analyzed. Results: A total of 300 embryos/spent media droplets from 54 patients aged 27-43 years (mean age ± SD: 36.33 ± 3.26) were evaluated. Of 111 embryos transferred, 19 implanted and led to a pregnancy: 7 (12.96%) single and 6 (11.1%) twin pregnancies. Irrespective of pregnancy, there were no systematic differences between the Raman spectra generated from spent media of Day 3 and Day 6 embryos, or between spent media and control media. Conclusions: In contrast to published reports, our study does not show that metabolomic profiling of spent embryo culture media by Raman spectroscopy can differentiate embryos with better implantation potential or add value to light microscopic assessment as in clinical practice.

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Attribution 3.0 Unported