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Non-invasive assessment of embryonic sex in cattle by metabolic fingerprinting of in vitro culture medium

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Abstract

The objective of this work was to determine whether metabolic fingerprinting of spent bovine embryo culture media using Fourier transform infrared spectroscopy (FTIR) correlates with embryonic sex. Embryos were produced in vitro from oocytes collected from cows slaughtered in an abattoir. Day-6 embryos were individually cultured in synthetic oviduct fluid for 24 h, prior to the time (Day-7) intended for embryo transfer or cryopreservation. Culture medium was analyzed by FTIR. Embryos were sexed by a PCR procedure based on amelogenin gene amplification or transferred to a recipient and sex observed at birth. Media samples from embryos diagnosed as male (n = 47) or female (n = 70) were individually collected and evaluated using FTIR. The spectra obtained were analyzed according to metabolomic profile of embryo culture media and embryonic sex. The discrimination capability of the classifiers was assessed for accuracy, sensitivity (female), sensitivity (male) and area under the ROC curve (AUC). Performance of sex prediction (%) was high within early blastocysts + blastocysts (74.4 ± 10.2, accuracy; 0.749 ± 0.099, AUC) and excellent for expanded blastocysts (86.0 ± 12.6, accuracy; 0.898 ± 0.094, AUC). A combination of metabolomic and bioinformatic analysis provides a non-invasive mean of embryonic sex analysis.

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Acknowledgments

Authors thank P. Bermejo-Alvarez and F. Goyache for valuable scientific comments, and JF Moreno (Sexing Technologies, Madison-Wisconsin, USA) for sexed semen donation. All institutional and national guidelines for the care and use of laboratory animals were followed.

Conflict of interest

MM, AU, EC, CD, AFG, JNC, BT, SC, ES and EG declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported.

Funding

This work was supported by the Spanish Ministry of Science and Innovation -MICINN– (AGL2009-10059). MM, EC and BT are supported by MICINN-RYC08-03454, MEC-FPU-AP2009-5265 and Cajastur, respectively. E.S. is supported by Award R01HD059909 from the National Institute of Health (NIH), USA. The authors are members of the COST Action FA1201 Epiconcept: Epigenetics and Periconception environment.

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Author notes

  1. Beatriz Trigal

    Present address: Sexing Technologies, 6938 Hickory Ln., DeForest, WI, 53532, USA

Authors and Affiliations

  1. Centro de Biotecnología Animal – SERIDA, Camino de Rioseco 1225, La Olla – Deva, 33394, Gijón, Asturias, Spain

    Marta Muñoz, Eva Correia, Carmen Díez, Jose Nestor Caamaño, Beatriz Trigal, Susana Carrocera & Enrique Gomez

  2. Department of Obstetrics, Gynecology, and Reproductive Sciences, Yale School of Medicine, 310 Cedar Street, LSOG 304D, New Haven, CT, 06520, USA

    Asli Uyar & Emre Seli

  3. Department of Computer Engineering, Okan University, Istanbul, Turkey

    Asli Uyar

  4. Servicios Científico Técnicos (Oviedo), Universidad de Oviedo, 33006, Oviedo, Asturias, Spain

    Alfonso Fernandez-Gonzalez

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  1. Marta Muñoz
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Corresponding author

Correspondence to Enrique Gomez.

Additional information

Marta Muñoz and Asli Uyar have contributed equally to this study.

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Muñoz, M., Uyar, A., Correia, E. et al. Non-invasive assessment of embryonic sex in cattle by metabolic fingerprinting of in vitro culture medium. Metabolomics 10, 443–451 (2014). https://doi.org/10.1007/s11306-013-0587-9

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