Non-invasive metabolomic profiling of day 3 and 5 embryo culture media using near infrared spectroscopy: predictions using pre-established models

Abstract

OBJECTIVE: ART pregnancy rates are high in the U.S., yet multiple embryo transfer continues as a treatment modality despite concerns regarding the incidence of multi-fetal pregnancies. Single embryo transfer (SET) is the only effective way to minimize the risks of multiple gestations. The objective of this study was to use a non-invasive metabolomic profiling technique based on Near Infra Red (NIR) spectroscopy to predict the implantation potential of Day 3 (D3) and 5 (D5) embryos that had been singly transferred.DESIGN: Analysis of embryo culture media from D3 and D5 embryos of SET cohorts in a private IVF center.MATERIALS AND METHODS: D3 and D5 SETs were performed at the Reproductive Science Center (RSC), Lexington, MA, USA (N=122 for D3 and N=25 for D5). Embryos were selected for SET using routine morphologic criteria and grading. Embryos were cultured individually in 30 μl under mineral oil to D3 in either Global or Sage Cleavage Medium and in Global Medium to D5. Post-SET, the medium in which the embryo had developed and a matching control medium were immediately frozen and sent to Molecular Biometrics, Inc. for analysis. Ongoing pregnancy was defined by the observation of fetal cardiac activity (FCA). A pre-established Molecular Biometrics D3 and D5 algorithm for a “Viability Index” predictive of pregnancy outcome was blindly assigned and applied to the embryo culture media samples collected at RSC.RESULTS: When the D3 embryo culutre media were blindly assessed, a significant difference (p=0.013) in the mean (±SD) “Viability Index” scores between the FCA- positive (0.51±0.23) and negative (0.39±0.31) samples was identified. A signficant (p=0.036) difference was also found for the smaller set of D5 SET media: FCA- positive (0.52±0.52) and negative (0.15±0.46). In general, in both data sets, as the Viability Index scores increased, their associated FCA-positive embryo implantations also increased.CONCLUSIONS: This study has shown that a pre-established D3 and 5 algorithm for analysis of spent embryo culture medium by NIR spectrophotometric analysis can contribute to the identification of embryos with a metabolomic profile, or “Viability Index” score, that may be predictive of implantation potential. Metabolomic profiling could provide a useful technology for assisting clinics in the US to move towards SET. OBJECTIVE: ART pregnancy rates are high in the U.S., yet multiple embryo transfer continues as a treatment modality despite concerns regarding the incidence of multi-fetal pregnancies. Single embryo transfer (SET) is the only effective way to minimize the risks of multiple gestations. The objective of this study was to use a non-invasive metabolomic profiling technique based on Near Infra Red (NIR) spectroscopy to predict the implantation potential of Day 3 (D3) and 5 (D5) embryos that had been singly transferred. DESIGN: Analysis of embryo culture media from D3 and D5 embryos of SET cohorts in a private IVF center. MATERIALS AND METHODS: D3 and D5 SETs were performed at the Reproductive Science Center (RSC), Lexington, MA, USA (N=122 for D3 and N=25 for D5). Embryos were selected for SET using routine morphologic criteria and grading. Embryos were cultured individually in 30 μl under mineral oil to D3 in either Global or Sage Cleavage Medium and in Global Medium to D5. Post-SET, the medium in which the embryo had developed and a matching control medium were immediately frozen and sent to Molecular Biometrics, Inc. for analysis. Ongoing pregnancy was defined by the observation of fetal cardiac activity (FCA). A pre-established Molecular Biometrics D3 and D5 algorithm for a “Viability Index” predictive of pregnancy outcome was blindly assigned and applied to the embryo culture media samples collected at RSC. RESULTS: When the D3 embryo culutre media were blindly assessed, a significant difference (p=0.013) in the mean (±SD) “Viability Index” scores between the FCA- positive (0.51±0.23) and negative (0.39±0.31) samples was identified. A signficant (p=0.036) difference was also found for the smaller set of D5 SET media: FCA- positive (0.52±0.52) and negative (0.15±0.46). In general, in both data sets, as the Viability Index scores increased, their associated FCA-positive embryo implantations also increased. CONCLUSIONS: This study has shown that a pre-established D3 and 5 algorithm for analysis of spent embryo culture medium by NIR spectrophotometric analysis can contribute to the identification of embryos with a metabolomic profile, or “Viability Index” score, that may be predictive of implantation potential. Metabolomic profiling could provide a useful technology for assisting clinics in the US to move towards SET.