OBJECTIVE: Embryo selection is based largely on morphology and is more difficult during SET. SET is the only effective way to stop the complication of multiple pregnancy. Non-invasive metabolomic profiling of embryo culture media using a NIR spectroscopy platform assesses modifications of the chemical composition of the embryo's medium and generates a Viability Score (VS). This score can rank each embryo's reproductive potential. The objective of this study was to assess the VS for individual patients' cohorts of Day 2 and 5 embryos and determine whether differences in scores exist in the embryo cohort.DESIGN: In clinic testing of embryo culture media.MATERIALS AND METHODS: From May-October 2009 a ViaMetrics-E™ Test Program was conducted in 13 IVF clinics in 4 countries. Embryos were cultured individually [20-40μl] and were selected for SET or multiple transfer based on morphology. After transfer up to 6 embryo media samples were assessed per patient, using the Viametrics-E embryo viability assessment procedure. In total, 672 Day 2 embryos from 220 cycles and 926 Day 5 embryos from 266 cycles were assessed. VS were generated for each embryo in the clinic.RESULTS: A mean of 3.1 Day 2 embryos and 3.5 Day 5 embryos were assessed for each patient. The mean (+ SD) VS was 0.49 + 0.3 for the Day 2 and 0.53 + 0.2 for the Day 5 embryos. When >3 embryos were available, the VS of the embryos within the cohort ranged betwen 0.1 to 0.9 for both Day 2 and Day 5. In the patients with >3 embryos, the highest scoring embryo(s) was selected for SET or multiple transfer in only 49/152 (32.2%) and 46/171 (26.9%) of cases for Day 2 and 5 respectively.CONCLUSION: It is evident that differences exist in the metabolomic state, as evidenced by their VS, in a given patients cohort of Day 2 or 5 embryos. This study supports the concept that only using morphology to assess embryos may fail to identify the embryo with the best metabolic and/or metabolomic profile within a cohort in the majority of transfers. OBJECTIVE: Embryo selection is based largely on morphology and is more difficult during SET. SET is the only effective way to stop the complication of multiple pregnancy. Non-invasive metabolomic profiling of embryo culture media using a NIR spectroscopy platform assesses modifications of the chemical composition of the embryo's medium and generates a Viability Score (VS). This score can rank each embryo's reproductive potential. The objective of this study was to assess the VS for individual patients' cohorts of Day 2 and 5 embryos and determine whether differences in scores exist in the embryo cohort. DESIGN: In clinic testing of embryo culture media. MATERIALS AND METHODS: From May-October 2009 a ViaMetrics-E™ Test Program was conducted in 13 IVF clinics in 4 countries. Embryos were cultured individually [20-40μl] and were selected for SET or multiple transfer based on morphology. After transfer up to 6 embryo media samples were assessed per patient, using the Viametrics-E embryo viability assessment procedure. In total, 672 Day 2 embryos from 220 cycles and 926 Day 5 embryos from 266 cycles were assessed. VS were generated for each embryo in the clinic. RESULTS: A mean of 3.1 Day 2 embryos and 3.5 Day 5 embryos were assessed for each patient. The mean (+ SD) VS was 0.49 + 0.3 for the Day 2 and 0.53 + 0.2 for the Day 5 embryos. When >3 embryos were available, the VS of the embryos within the cohort ranged betwen 0.1 to 0.9 for both Day 2 and Day 5. In the patients with >3 embryos, the highest scoring embryo(s) was selected for SET or multiple transfer in only 49/152 (32.2%) and 46/171 (26.9%) of cases for Day 2 and 5 respectively. CONCLUSION: It is evident that differences exist in the metabolomic state, as evidenced by their VS, in a given patients cohort of Day 2 or 5 embryos. This study supports the concept that only using morphology to assess embryos may fail to identify the embryo with the best metabolic and/or metabolomic profile within a cohort in the majority of transfers.

OBJECTIVE: Non-invasive profiling of embryo culture media using NIR spectroscopy assesses changes of the chemical composition of the embryo's surrounding medium to provide information about the potential viability of embryos. The objective of this study was to assess the media from Day 2 and 5 embryos and examine whether a NIR spectroscopy procedure [ViaMetrics E™] could assist in deciding which embryo to transfer on the same day. DESIGN: In clinic testing of embryo culture media. MATERIALS AND METHODS: From May to October 2009 a ViaMetrics-E Test Program was conducted in 13 IVF clinics in 4 countries. Embryos were cultured individually [20-40μl] and selected for transfer using morphology. After transfer, up to 6 embryo media samples were assessed per patient using the embryo viability assessment procedure. Over 2400 spectra were collected and analyzed, including 539 transferred embryos. Analysis was performed to examine: (i) the quality of the spectra and (ii) the clinical outcomes associated with the spectra and their subsequent Viability Scores. Clinical outcome was measured when the embryo(s) transfer number equaled Fetal Heart Beats (ie. 100% implantation rate). RESULTS: Due to the introduction of air bubbles prior to assessment of media, 12.5% (range 4-25% per clinic) of embryo transfer samples could not be analyzed. Of the remaining spectra, 175 Day 2 and 189 Day 5 transfers led to a 100% implantation rate, ie. the Viability Score could be correlated to implantation potential of individual embryos. For both transfer days a positive correlation was observed between an increasing Viability Score and increasing implantation rate [Day 2: R=0.77 and Day 5: R=0.98]. CONCLUSION: As a result of this study, procedural changes have been made to improve spectral quality and eliminate air bubbles. Importantly, NIR assessment of the culture media in the clinic appears to generate Viability Scores that constitute a parameter, in addition to morphology, for a rapid non-invasive assessment of embryo viability.

OBJECTIVE: Current embryo assessment strategies rely largely on morphology and cleavage rate in order to identify embryos that are most likely to implant in women undergoing in vitro fertilization (IVF). In this study, we assessed whether a viability score generated by biospectroscopy-based metabolomic profiling of spent embryo culture media correlates with implantation potential of individual embryos in women undergoing single embryo transfer (SET) on day 2.DESIGN: A prospective multi-center, multi-national study was conducted in 4 IVF centers.MATERIALS AND METHODS: A total of 176 spent embryo culture media samples were collected following single embryo transfer (SET) on day 2 at Unilabs Laboratoire D'Eylau, Paris, France; Vaestoliitto Fertility Clinics, Helsinki, Finland; Fertilitetscentrum, Gothenburg, Sweden; and Royal Women's Hospital, Melbourne, Australia. Samples were analyzed by near infrared (NIR) spectroscopy using a wavelength-specific genetic algorithm and a relative embryo viability score was generated for each sample. All measurements were performed at Molecular Biometrics Inc. by observers blinded to embryo morphology or IVF outcome. Pregnancy was defined as fetal cardiac activity (FCA) at 12 weeks of gestation.RESULTS: Viability scores of embryos that resulted in pregnancy (FCA+) were significantly higher compared to embryos that failed to implant (FCA–) (mean±SD; 0.600±0.182 vs. 0.529±0.172; p=0.011). A positive correlation was observed between increasing viability index values and the reproductive potential of individual embryos (Pearson correlation coefficient 0.18; p<0.05). There was not a signifcant correlation between viability scores and embryo morphology or type of culture media used.CONCLUSIONS: Viability scores determined by metabolomic profiling of day 2 spent embryo culture media correlate with outcome in women undergoing IVF and constitute a parameter independent of morphology for non-invasive assessment of embryo viability. OBJECTIVE: Current embryo assessment strategies rely largely on morphology and cleavage rate in order to identify embryos that are most likely to implant in women undergoing in vitro fertilization (IVF). In this study, we assessed whether a viability score generated by biospectroscopy-based metabolomic profiling of spent embryo culture media correlates with implantation potential of individual embryos in women undergoing single embryo transfer (SET) on day 2. DESIGN: A prospective multi-center, multi-national study was conducted in 4 IVF centers. MATERIALS AND METHODS: A total of 176 spent embryo culture media samples were collected following single embryo transfer (SET) on day 2 at Unilabs Laboratoire D'Eylau, Paris, France; Vaestoliitto Fertility Clinics, Helsinki, Finland; Fertilitetscentrum, Gothenburg, Sweden; and Royal Women's Hospital, Melbourne, Australia. Samples were analyzed by near infrared (NIR) spectroscopy using a wavelength-specific genetic algorithm and a relative embryo viability score was generated for each sample. All measurements were performed at Molecular Biometrics Inc. by observers blinded to embryo morphology or IVF outcome. Pregnancy was defined as fetal cardiac activity (FCA) at 12 weeks of gestation. RESULTS: Viability scores of embryos that resulted in pregnancy (FCA+) were significantly higher compared to embryos that failed to implant (FCA–) (mean±SD; 0.600±0.182 vs. 0.529±0.172; p=0.011). A positive correlation was observed between increasing viability index values and the reproductive potential of individual embryos (Pearson correlation coefficient 0.18; p<0.05). There was not a signifcant correlation between viability scores and embryo morphology or type of culture media used. CONCLUSIONS: Viability scores determined by metabolomic profiling of day 2 spent embryo culture media correlate with outcome in women undergoing IVF and constitute a parameter independent of morphology for non-invasive assessment of embryo viability.

OBJECTIVE: The objective of this study was to use a non-invasive metabolomic profiling technique, based on NIR Spectroscopy, to blindly predict the reproductive potential of Day 2 embryos that had undergone SET. DESIGN: Analysis of embryo culture media from day 2 embryos that had undergone SET in a private IVF center. MATERIALS AND METHODS: Day 2 SETs [N=45] were performed at the Väestöliitto Fertility Clinic (VFF) , Helsinki, Finland. Embryos had been cultured individually in Vitrolife G-1 Medium in 25μl droplets under oil and selected for SET based on morphology. After SET, the medium in which the embryo was cultured and a control medium were frozen. The stored media were then sent to Molecular Biometrics for analysis. Pregnancy was defined by assessing positive fetal cardiac activity (FCA). A Day 2 algorithm has been previously developed by examining the NIR spectra of positive and negative FCA outcomes. A relative “Viability Score”, predictive of pregnancy outcome, was assigned blindly by the algorithm to embryo culture media samples collected at VFF. RESULTS: When the transferred media were assessed we found a significant difference (P=0.002) in the mean Viability Scores between the FCA negative and positive samples. As the Viability Scores increase their associated positive FCA rates also increase, reaching an R squared value of 0.93.The results are shown in Table 1.Table 1Blinded Prediction of FCA outcome in relation to increasing Viability Scores.Viability ScoreNo. of FCA PositiveNo. of FCA Negative% Positive FCA<0.21109.10.2-0.332918.20.33-0.55645.5>0.56650.0 Open table in a new tab CONCLUSIONS: This study has shown that a pre-established Day 2 algorithm significantly improves the probability of predicting the viability of an individual embryo. The blinded prediction of the Day 2 media samples indicates that the viability score can be used as a rapid non-invasive addition to embryo morphology in predicting pregnancy prior to SET.

OBJECTIVE: Non-invasive metabolomic profiling of embryo culture media using NIR spectroscopy assesses modifications of the chemical composition of the embryo's surrounding medium and generates a Viability Score. The objective of this study was to assess the Viability Score for an individual patient's cohort of similar good grade morphological embryos and determine whether differences in scores could impact on the patient's chances of achieving pregnancy.DESIGN: Analysis of day 2 embryo culture media.MATERIALS AND METHODS: Embryos were cultured individually in 20μl of SAGE cleavage medium at the Kato Ladies Clinic, Tokyo, Japan. Media samples were collected from 95 good grade 1 and 2 morphology embryos that were selected for either transfer or freezing from 36 patients. The embryo media samples and blank controls were frozen and stored. NIR spectral analysis was performed on all the embryo culture media samples and expressed as a Viability Score. The score reflects the implantation potential of an embryo.RESULTS: On average (mean + SD) 2.6 + 1.2 good grade embryos were assessed for each patient. NIR analysis of all 95 embryo media gave varying scores ranging from 0 to 0.6. Within an individual patient's cohort of embryos the average variability in the Viability Score was 0.34 + 0.1. We have previously shown that Day 2 Viability Scores of <0.3 or >0.3 give an implantation rate of 20% and 41% respectively (Seli et al. Fertil. Steril. 2009). The selection of a good morphology embryo with a Viability Score of 0.24 would have an expected implantation potential of ∼20% while a similar morphology embryo scoring 0.44 would have an expected implantation potential of ∼40%.CONCLUSIONS: The current study shows that the Viability Score varies significantly within a patient's cohort of good embryos. This indicates that assessing a metabolic parameter could have an enormous impact on improving the chances of selecting the most viable embryo within a cohort of morphologically good grade embryos. OBJECTIVE: Non-invasive metabolomic profiling of embryo culture media using NIR spectroscopy assesses modifications of the chemical composition of the embryo's surrounding medium and generates a Viability Score. The objective of this study was to assess the Viability Score for an individual patient's cohort of similar good grade morphological embryos and determine whether differences in scores could impact on the patient's chances of achieving pregnancy. DESIGN: Analysis of day 2 embryo culture media. MATERIALS AND METHODS: Embryos were cultured individually in 20μl of SAGE cleavage medium at the Kato Ladies Clinic, Tokyo, Japan. Media samples were collected from 95 good grade 1 and 2 morphology embryos that were selected for either transfer or freezing from 36 patients. The embryo media samples and blank controls were frozen and stored. NIR spectral analysis was performed on all the embryo culture media samples and expressed as a Viability Score. The score reflects the implantation potential of an embryo. RESULTS: On average (mean + SD) 2.6 + 1.2 good grade embryos were assessed for each patient. NIR analysis of all 95 embryo media gave varying scores ranging from 0 to 0.6. Within an individual patient's cohort of embryos the average variability in the Viability Score was 0.34 + 0.1. We have previously shown that Day 2 Viability Scores of <0.3 or >0.3 give an implantation rate of 20% and 41% respectively (Seli et al. Fertil. Steril. 2009). The selection of a good morphology embryo with a Viability Score of 0.24 would have an expected implantation potential of ∼20% while a similar morphology embryo scoring 0.44 would have an expected implantation potential of ∼40%. CONCLUSIONS: The current study shows that the Viability Score varies significantly within a patient's cohort of good embryos. This indicates that assessing a metabolic parameter could have an enormous impact on improving the chances of selecting the most viable embryo within a cohort of morphologically good grade embryos.

OBJECTIVE: We have recently reported that non-invasive metabolomic profiling of embryo culture media using near infrared (NIR) spectroscopy correlates with the reproductive potential of embryos in IVF (Seli et al., 2007). This technology assesses modifications of the chemical composition of the embryo's surrounding medium using spectroscopy and generates a value termed the "ViaTest-E™ score". In the current study, we hypothesized that the use of a ViaTest-E score could further assist in determining the most viable embryo in addition to extended culture to blastocyst. DESIGN: NIR analysis of Day 5 embryo culture media samples and correlation with pregnancy outcome. MATERIALS AND METHODS: Single embryo Day 5 transfers (n=137) were performed at the Fertilitetscentrum, Gothenburg, Sweden and Shady Grove Fertility RSC, Rockville, Maryland, USA. Day 5 embryos were assessed for SET based on morphology. After SET a media sample from the transferred embryo and a blank were collected and cryostored. Media NIR analysis and a genetic algorithm were used to identify spectral regions, for eg. -SH, C=C, -CH, -OH, and –NH groups, that discriminated most between transferred embryos that did and did not result in pregnancy (fetal cardiac activity detected at 12 weeks of gestation). Relative ViaTest-E scores were calculated by quantification of these spectral regions, adjusting for parallel blank media controls. RESULTS: The overall pregnancy rate from the two clinical sites using SET was 69/137 (50.4%). When the transferred embryo's ViaTest-E scores were assessed, we found a large variability in metabolic profiles (range: -0.5 to 1.5). A significant (Pearsons: P<0.001) positive correlation was observed with increasing ViaTest-E score quintiles and their associated implantation rates (Table 1). CONCLUSIONS: The ViaTest-E score helps discriminate between day 5 embryos, confirming that the embryo's metabolism varies widely regardless of morphology. The added armory of metabolomic profiling by NIR spectroscopy, coupled with bioinformatics, to current morphological assessment protocols seems to allow a greater discrimination for selection of embryos for transfer, and has considerable promise to improve IVF outcomes.Table 1ViaTest-E scores of Day 5 embryos and implantation ratesViaTest-E scorePregnantNot PregnantImplantation Rate (%)<0.252271926.90.2522-0.445162122.20.4451-0.5415131448.10.5415-0.700421677.8>0.700422873.3 Open table in a new tab

BACKGROUNDThe morphology of an embryo has a limited predictive value for assessing viability and ongoing pregnancy, therefore new selection tools are needed to maintain success rates with single-embryo transfer (SET). In this study, we investigated if metabolomic profiling of biomarkers of embryo culture medium by near-infrared (NIR) spectroscopy has a correlation with ongoing pregnancy in SET.METHODSA total of 333 patients scheduled for in vitro fertilization (IVF) with SET were included in the study. Embryos were selected for transfer by morphological criteria on Days 2 and 3 of in vitro culture, and left over culture media samples were analyzed by NIR spectroscopy.RESULTSThe NIR spectral analysis produced unique metabolomic profiles that correlated to an embryo's reproductive potential. Resulting relative viability scores between positive and negative pregnancy outcomes were statistically significant (P < 0.03). A logistic regression of factors correlated to pregnancy outcomes showed that maternal age, percent fragmentation and relative viability scores all demonstrated a relationship. The extent of the correlation was determined by accuracy computation, where the accuracy of assessing viable embryos on Day 3 by metabolomic profiling was 53.6% and the accuracy of the morphological selection was 38.5%. In addition, the positive predictive value of metabolomic profiling was 0.365 and the negative predictive value was 0.830.CONCLUSIONSNIR metabolomic profiling of spent embryo culture media was able to distinguish viable embryos from non-viable embryos for reproduction.

To assess status of metabolic activity and viability of D2 embryos selected for SET using non-invasive metabolomic profiling of biomarkers of oxidative metabolism (OM). Prospective, blinded study. A “mild” ovarian stimulation protocol using 50 mg clomiphene citrate daily combined with 50IU hMG on day 8 was used until GnRHa administration prior to OPU. Fertilized zygotes were cultured individually in 20ul of cleavage medium (SAGE) with 12 mg/ml SPS. Discarded media samples were collected, frozen and stored in liquid N2 on D2. Control media samples (CMS) were obtained for each embryo media specimen collected. Near Infrared spectroscopy (NIR) measurements of blinded, randomized samples were conducted using an InGaAs spectrometer; sample volume = 7 ul. CMS were used to compensate for signal drift. Sample properties were quantified from resulting mean centered NIR spectra by determining discrete variables in selected wavelength (WL) domains using genetic algorithm optimization. Selected WL regions were weighted by a coefficients calculated by inverse least-squares regression. Each sample's pregnancy viability was estimated by a leave-one-out cross validation method. Notch box plots were used to plot resulting viability indices; t-tests were applied to determine statistical significance. 159 D2 media specimens and 159 CMS were analyzed. NIR spectral analysis reproducibly defined 5 to 6 OM biomarker regions associated with changes in ROM, C=C, -SH, -CH and -NH functional groups that were used to distinguish between FCA+ vs. FCA- using proprietary bioinformatics. The FCA+/FCA− groups were statistically different at P≤.05. Morphology scores of embryos between FCA+ and FCA− groups were not statistically different indicating no correlation with prediction of reproductive potential or metabolomic activity. Studies implicate OM as a factor affecting embryo development and viability. D2 is a crucial stage of embryo growth and development involving activation of new metabolic pathways. Unlike metabolomic data obtained by NIR, morphological criteria were not correlated with embryo competency. Variations in drugs and dosages used in different stimulation protocols may affect metabolomic activity and quality in developing (D2) embryos (data on file). Metabolomic profiling appears to be a reliable indicator of metabolic activity and embryo viability as early as D2, whereas morphology was not predictive of either parameter.

SET is an effective method to minimize risks of multiple pregnancies in IVF. Since morphology has limited predictive value, new selection tools are needed to ensure success rates are maintained. Previous studies have shown that non-invasive MetPro may be useful in predicting reproductive potential of embryos. The purpose of this study was to investigate the efficiency of MetPro of biomarkers of OM in patients undergoing SET compared to morphological selection. Prospective study. 274 patients scheduled for IVF with SET received standard gonadotropin stimulation protocols with agonist down-regulation. Embryos were cultured individually in 25 μl media drops (HTF, Cambrex, Belgium and 10% GPO) alongside embryo-free media drops as controls. Embryos were selected for transfer by strict morphological criteria. After transfer, spent media were immediately frozen (−196°C). Pregnancy was defined by positive fetal cardiac activity (FCA) at 12 weeks. 7 μl media samples were analysed by Near Infrared (NIR) spectroscopy; analysis time was <1 min. OM biomarkers of ROH, -SH, C = C, -CH, -OH, and -NH groups were identified yielding unique metabolomic profiles that were quantified using a wavelength selective genetic algorithm, proprietary bioinformatics and leave-one out cross-validation, in conjunction with logistical regression, to create a relative “embryo viability score” that correlated to pregnancy outcomes. The resulting metabolomics data were compared to morphological assesment and pregnancy outcomes. NIR spectral analysis of discarded culture media samples produced unique metabolomic profiles of OM that correlated to an embryo's reproductive potential as determined by FCA. Differences in metabolomic profiles between FCA+ and FCA− groups were statistically significant (>95% confidence interval); sensitivity and specificity was 87% and 93%, respectively. Greater than 86% of all embryos transferred were classified as Grade A or B embryos by strict morphological criteria while 27% resulted in an ongoing pregnancy. Conversely, significant numbers of poorer quality embryos (Grade C and D) actually resulted in FCA+ outcomes. MetPro of discarded embryo culture media by NIR was effective in distinguishing between viable and non-viable embryos in SET irrespective of embryo morphology. MetPro targeting biomarkers of OM may provide a strong addition for rapid, non-invasive selection of embryos with high reproductive potential in SET.

MetPro of biomarkers of OM has been used successfully to identify the reproductive potential of embryos following IVF and multiple ET. In this study we applied metabolomics, based on Near Infrared (NIR) spectroscopy, to ascertain the reproductive potential of day-5 embryos in SET. Prospective, blinded. Infertile women (n = 23) and oocyte donors (n = 11) were down-regulated and stimulated with gonadotropins. IVF embryos were cultured individually in 80 μl drops of sequential media. Embryos were graded and selected for SET on day-5 based on morphology. Following ET 25 μl of spent-medium was collected and stored at −80°C. Pregnancy was determined by ultrasound detection of fetal cardiac activity (FCA) at 6.5 weeks. NIR measurements of samples were conducted using an InGaAs spectrometer (vol. = 7 μl; analysis time <1 min). Media controls were used to compensate for signal drift. Sample properties were quantified from the resulting mean centered NIR spectra by determining the most parsimonious combination of variables in selected wavelength domains using genetic algorithm optimization. Selected wavelength regions were weighted by coefficients calculated by inverse least-squares regression. Each sample's pregnancy viability was estimated in a continuous reproductive potential index by a leave-one-out cross validation method. Notch box plots were used to plot resulting viability indices, and statistical significances were determined by t-tests. Overall pregnancy rate was 56%. NIR spectral analysis reproducibly defined 4 to 5 OM biomarker regions associated with changes in ROH, C=C, -SH, -CH and -NH functional groups that were used to distinguish a statistically significant difference between FCA+ vs. FCA− samples. Metabolomic profiles of donor embryos showed greater differences between FCA+ and FCA− results than did profiles of embryos produced by infertile women; the latter had considerably more variation in OM in the FCA+ group. Morphology score did not correlate with pregnancy outcome. In this SET study, non-invasive MetPro revealed statistical differences between all FCA+ embryos compared to all FCA- embryos. FCA+ embryos of infertile women were more variable and had lower predictive scores than those of donors. Morphology was not correlated with embryo implantation. MetPro appears to offer an exciting new paradigm for selecting embryos with the greatest reproductive potential, beyond conventional morphological grading, for optimization of day-5 SET.

It has been recently proposed that biospectroscopy-based metabolomics can be used to assess biomarkers of oxidative metabolism (OM), from culture medium of embryos. Our objective was to evaluate, if it is possible to establish novel metabolomic data from culture medium obtained from different maturational stage oocytes. Furthermore, we wanted to assess how metabolomic profiles, obtained for each group of maturational stage oocytes, correlate with levels of oxidative metabolism. Prospective collection and evaluation of oocyte culture medium by infrared spectroscopy. ICSI patients with more than 10 eggs and under age of 37 were included. After three hours of incubation of eggs in individual drops, oocyte media was collected representing the different maturation stages. Selected embryos were transferred on Day 3 or 5. Individual profiles were obtained from 5 μl media samples using Near Infrared (NIR) spectroscopy. Specific biomarkers, corresponding to unique functional groups (NH2, aromatic -CH, -OH, -SH) were identified yielding exclusive metabolomic profiles which were then quantified using a wavelength selective algorithm (Molecular Biometrics, LLC, Chester, NJ). Resulting metabolomics data were correlated with morphological assessed nuclear maturity of the corresponding oocytes using multivariate analyses with a P level of 5%. A total of 264 oocyte culture samples were collected from 32 patient cycles. Unique metabolomic profiles of the specific OM biomarker populations were consistently identified in all samples by NIR spectroscopy. Profiles of Metaphase I and II oocytes were significantly different from each other and from profiles of Prophase I (GV) oocytes (P<0.001 at the 95% confidence interval). NIR analysis of biomarkers required a minimum of three wavelength regions, and resulted in a specificity of 83% and a sensitivity of 85% for GV vs. MI oocytes; and 93% specificity and 100% sensitivity between MI and MII oocytes. The results of this study demonstrate that a detectable and significant difference exists in the metabolomic profiles of OM biomarkers found in culture media obtained from oocytes with the different maturational stages. These data confirm that NIR analyses using can serve to assess metabolomic status also at the gamete level with high sensitivity (only 3 h of incubation). Further studies are planned to evaluate if gamete metabolomic profiling can also correlate with corresponding embryo development.