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

Eldarov, Chupalav; Gamisonia, Alina M.; Chagovets, Vitaliy; Luiza, Ibragimova; Yarigina, S A; Smolnikova, Veronika; Kalinina, Kalinina E.A.; Макарова, Н. П.; Zgoda, Victor G.; Сухих, Г. Т.; Bobrov, M. Yu.

doi:10.3390/ijms23052706

Cited by 8 publications

(10 citation statements)

References 80 publications

(83 reference statements)

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“…A similar profile was found for human embryos, in which linoleic acid metabolism and steroid biosynthesis were indicated as enriched pathways of molecular ions from spent culture media that successfully or unsuccessfully establish implantation (Eldarov et al, 2022).…”

Section: Ta B L E 1 Upregulated Pathways In Group Pregnant Pathways A...supporting

confidence: 64%

“…Stearic and Capric acid, with decreased embryo viability and poor pregnancy outcomes (Gomez et al, 2021). A similar profile was found for human embryos, in which linoleic acid metabolism and steroid biosynthesis were indicated as enriched pathways of molecular ions from spent culture media that successfully or unsuccessfully establish implantation (Eldarov et al, 2022). Indeed, high saturated FA concentrations have been shown to be detrimental for oocyte development and embryo quality (Desmet et al, 2016, 2020).…”

Section: Discussionsupporting

confidence: 53%

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Metabolic signature of spent culture media shows lipid metabolism as a determinant of pregnancy outcomes

Fernandes

Lima

Fidelis

et al. 2022

Reprod Domestic Animals

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In the present study, we investigated the spent culture media of in vitro produced (IVP) bovine embryos which did (group Pregnant) or did not (group Non-pregnant) establish pregnancy after transfer. For that purpose, IVP embryos on D5 were transferred to individual droplets for the last 48 h of culture. Embryos at the blastocyst stage were then transferred to synchronized recipients, while respective culture media drops were collected and evaluated individually. The list of metabolites present in spent culture media was obtained by electrospray ionization mass spectrometry (ESI-MS) and analysed with Metaboanalyst® to characterize the metabolic profile of each group. The spectrometric analysis showed that pathways related to lipid metabolism, particularly fatty acids degradation via beta-oxidation, were more present in the Pregnant group whereas no significant pathway was identified in the group Non-pregnant. By using this method, we were able to identify a metabolic signature in culture media that allows for a better comprehension of preferential metabolic routes taken by the most viable embryos. These findings offer great insights into the biochemistry of embryo development and reveal a potential target for the development of better-quality IVP systems, as well as tools to identify bovine embryos with greater chances to establish and maintain pregnancy.

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Section: Ta B L E 1 Upregulated Pathways In Group Pregnant Pathways A...supporting

confidence: 64%

Section: Discussionsupporting

confidence: 53%

Metabolic signature of spent culture media shows lipid metabolism as a determinant of pregnancy outcomes

Fernandes

Lima

Fidelis

et al. 2022

Reprod Domestic Animals

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“…A reliable noninvasive method to choose the best candidate with real implantation potential for transfer from two embryos with equal morphological qualities is still lacking clinically, which hampered the improvement of single embryo transfer stratege. Spent embryo culture medium could be collected noninvasively after embryo was cultivated, and its metabolomic pro ling relevent with embryo implantation potential could be detected by a range of spectroscopic methods including Fourier Transform infrared spectroscopy (FTIR), near-infrared spectroscopy (NIR), Raman spectroscopy [7,8], NMR [9] and liquid chromatography Mass-spectrometry (LC-MS) [10,11]. Then an approach basing on one of these metabolomic pro ling detection methods provids a potential adjunct to embryo morphological assessment.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, high sensitive metabolomic pro ling detection method such as LC-MS may provide more information. In 2022, Eldarov C et al performed HPLC-MS on SECM samples from day 3 embryos of the rst morphological class with corresponding transfer outcomes and they selected 50 molecular irons with a signi cant fold change between the s-implanting and f-implanting group [11]. They concluded HPLC-MS for SECM metabolomic analysis could be a powerful and sensitive tool for embryo implantation potential.…”

Section: Discussionmentioning

confidence: 99%

LC-MS detected universal metabolites of human day 3 embryos with distinct implantation outcomes for Cook Medical and Vitrolife culture medium

Shi

Liu

et al. 2022

Preprint

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Background To implore the universal metabolites from metabolomic profiling of spent embryo culture medium (SECM) correlated to day 3 embryo’s implantation potential in two commercial culture medium (Cook Medical and Vitrolife). Methods This investigation was a prospective randomized study. Patients undergoing IVF-ET and having utilizable day 3 embryos were recruited. Patients were randomized for embryo culture in Cook Medical or Vitrolife medium. On day 1 each zygote of patients was individually cultured in a 30 µL microdroplet until day 3. Then SECM from each microdroplet was collected within 2 hours after the embryo was utilized (transferred or frozen) and detected by Liquid Chromatography-Mass Spectrometry (LC-MS). Results For Vitrolife medium, 3 batches of total 69 SECM samples were collected. According to the embryo’s implantation outcome, 17 were successfully implanted group, while 52 were failed implanted group. 147 metabolomic irons presenting significantly different level between the two groups were identified as the Vitrolife metabolomic profiling set. For Cook Medical medium, 2 batches of total 37 SECM samples were collected. According to the embryo’s implantation outcome, 14 were successfully implanted group, while 23 were failed implanted group. 435 metabolomic irons presenting significantly different level between the two groups as the Cook Medical metabolomic profiling set. There were 66 universal metabolites between the Vitrolife and Cook Medical metabolomic profiling set and the ion with m/z = 121.029 was the most reproduciable signature, which was identified to be benzoic acid. Conclusions Metabolomic profiling in SECM correlated to day 3 embryo’s implantation potential in Vitrolife and Cook Medical medium could be sensitively detected by LC-MS approach. The universal metabolites for the two commercial medium, such as benzoic acid, may provide a potential solid adjunct to day 3 embryo’s evaluation.

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“…Interestingly, a comparative analysis of the metabolomic profiles of SECM on day 5 found two different clusters of metabolite composition between euploid and aneuploid embryos with good morphology. Fur-thermore, untargeted metabolomics of SECM by high-performance liquid chromatography-mass spectrometry identified potential biomarkers of embryos with good morphology that would undergo unsuccessful implantation [83]. In a preliminary report, three artificial neural networks that combined morphological variables and proteins using blastocyst image analysis and proteomic profiles of SECM were able to predict live birth, with an AUC of 1.0 in receiver operating characteristic curve analysis [84].…”

Section: Development and Advancement Of Molecular Biological Methods ...mentioning

confidence: 99%

Non-invasive evaluation of embryo quality for the selection of transferable embryos in human in vitro fertilization-embryo transfer

Kim

Lee

Jun

2022

Clin Exp Reprod Med

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The ultimate goal of human assisted reproductive technology is to achieve a healthy pregnancy and birth, ideally from the selection and transfer of a single competent embryo. Recently, techniques for efficiently evaluating the state and quality of preimplantation embryos using time-lapse imaging systems have been applied. Artificial intelligence programs based on deep learning technology and big data analysis of time-lapse monitoring system during in vitro culture of preimplantation embryos have also been rapidly developed. In addition, several molecular markers of the secretome have been successfully analyzed in spent embryo culture media, which could easily be obtained during in vitro embryo culture. It is also possible to analyze small amounts of cell-free nucleic acids, mitochondrial nucleic acids, miRNA, and long non-coding RNA derived from embryos using real-time polymerase chain reaction (PCR) or digital PCR, as well as next-generation sequencing. Various efforts are being made to use non-invasive evaluation of embryo quality (NiEEQ) to select the embryo with the best developmental competence. However, each NiEEQ method has some limitations that should be evaluated case by case. Therefore, an integrated analysis strategy fusing several NiEEQ methods should be urgently developed and confirmed by proper clinical trials.

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

Cited by 8 publications

References 80 publications

Metabolic signature of spent culture media shows lipid metabolism as a determinant of pregnancy outcomes

Metabolic signature of spent culture media shows lipid metabolism as a determinant of pregnancy outcomes

LC-MS detected universal metabolites of human day 3 embryos with distinct implantation outcomes for Cook Medical and Vitrolife culture medium

Non-invasive evaluation of embryo quality for the selection of transferable embryos in human in vitro fertilization-embryo transfer

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