1H NMR based profiling of spent culture media cannot predict success of implantation for day 3 human embryos
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Identification of a non-invasive technique to assess embryo implantation potential in assisted reproduction would greatly increase success rates and lead more efficiently to single embryo transfer. Early studies suggested metabonomic analysis of spent culture media could improve embryo selection. The goal of this study is to assess if embryo implantation can be predicted based on proton nuclear magnetic resonance (1H NMR) profiles of spent embryo culture media from patients undergoing transfer of multiple embryos on cycle day 3.
We conducted a retrospective study in an academic assisted reproduction technology (ART) program and analyzed the data in a university research center. Two hundred twenty-eight spent culture media samples originating from 108 patients were individually analyzed. Specifically, five distinct sets (1 to 5) of different types of spent media samples (volume ~14 μL) from embryos that resulted in clinical pregnancy (positive heart rate at 6 weeks gestation) (n 1 = 29; n 2 = 19; n 3 = 9; n 4 = 12; n 5 = 33; n total = 102) and from embryos that did not implant (n 1 = 28; n 2 = 29; n 3 = 18; n 4 = 15; n 5 = 36; n total = 126) were collected on day 3 of embryo growth. The media samples were profiled using 1H NMR spectroscopy, and the NMR profiles of sets 1 to 5 were subject to standard uni- and multi-variate data analyses in order to evaluate potential correlation of profiles with implantation success.
For set 1 of the media samples, a borderline class separation of NMR profiles was obtained by use of principal component analysis (PCA) and logistic regression. This tentative class separation could not be repeated and validated in any of the other media sets 2 to 5.
Despite the rigorous technical approach, 1H NMR based profiling of spent culture media cannot predict success of implantation for day 3 human embryos.
KeywordsART 1H NMR Metabonomics IVF Implantation success
- 1.2009 Guidelines on number of embryos transferred. Fertil Steril. 92 1518–9.Google Scholar
- 3.Benjamini Y, Hochberg Y. Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc Ser B (Methodological). 1995;57:289–300.Google Scholar
- 4.Blake DA, Farquhar CM, Johnson N, Proctor M. Cleavage stage versus blastocyst stage embryo transfer in assisted conception. Cochrane Database Syst Rev. 2007;CD002118.Google Scholar
- 15.Lindon JC, Nicholson JK, Holmes E. The Handbook of metabonomics and metabolomics. Amsterdam: Elsevier; 2007.Google Scholar
- 18.Pudakalakatti SM, Uppangala S, D’Souza F, Kalthur G, Kumar P, Adiga SK, Atreya HS. NMR studies of preimplantation embryo metabolism in human assisted reproductive techniques: a new biomarker for assessment of embryo implantation potential. NMR Biomed. 2012Google Scholar
- 27.Seli E, Bruce C, Botros L, Henson M, Roos P, Judge K, Hardarson T, Ahlstrom A, Harrison P, Henman M, Go K, Acevedo N, Siques J, Tucker M, Sakkas D. Receiver operating characteristic (ROC) analysis of day 5 morphology grading and metabolomic Viability Score on predicting implantation outcome. J Assist Reprod Genet. 2011;28:137–44.PubMedCrossRefGoogle Scholar
- 28.Seli E, Sakkas D, Scott R, Kwok SC, Rosendahl SM, Burns DH. Noninvasive metabolomic profiling of embryo culture media using Raman and near-infrared spectroscopy correlates with reproductive potential of embryos in women undergoing in vitro fertilization. Fertil Steril. 2007;88:1350–7.PubMedCrossRefGoogle Scholar
- 33.Vergouw CG, Kieslinger DC, Kostelijk EH, Hompes PG, Schats R & Lambalk CB. Metabolomic profiling of culture media by near infrared spectroscopy as an adjunct to morphology for selection of a single day 3 embryo to transfer in Ivf: a double blind randomised trial. In American Society of Reproductive Medicine O:7, 2011.Google Scholar