In vitro culture and non-invasive metabolic profiling of single bovine embryos
Monika Nõmm
A F , Rando Porosk B D , Pille Pärn A C , Kalle Kilk B D , Ursel Soomets B D , Sulev Kõks A E and Ülle Jaakma A C
+ Author Affiliations
- Author Affiliations
A Chair of Animal Breeding and Biotechnology, Institute of Veterinary Medicine and Animal Sciences, Estonian University of Life Sciences, Fr. R. Kreutzwaldi 1, Tartu 51006, Estonia.
B Department of Biochemistry, Institute of Biomedicine and Translational Medicine, University of Tartu, Ülikooli 18, Tartu 50090, Estonia.
C Competence Centre on Health Technologies, Tiigi 61b, Tartu 50410, Estonia.
D Centre of Excellence for Genomics and Translational Medicine, University of Tartu, Ülikooli 18, Tartu 50090, Estonia.
E Department of Pathophysiology, Institute of Biomedicine and Translational Medicine, University of Tartu, Ülikooli 18, Tartu 50090, Tartu, Estonia.
F Corresponding author. Email: monika.nomm@emu.ee
Reproduction, Fertility and Development 31(2) 306-314 https://doi.org/10.1071/RD17446
Submitted: 24 October 2017 Accepted: 29 June 2018 Published: 10 August 2018
Abstract
Selecting high-quality embryos for transfer has been a difficult task when producing bovine embryos in vitro. The most used non-invasive method is based on visual observation. Molecular characterisation of embryo growth media has been proposed as a complementary method. In this study we demonstrate a culture medium sampling method for identifying potential embryonic viability markers to predict normal or abnormal embryonic development. During single embryo culture, 20 µL culture media was removed at Days 2, 5 and 8 after fertilisation from the same droplet (60 µL). In all, 58 samples were analysed using liquid chromatography–mass spectrometry. We demonstrate that it is possible to remove samples from the same culture medium droplets and not significantly affect blastocyst rate (25.2%). Changes in any single low molecular weight compound were not predictive enough. Combining multiple low molecular weight signals made it possible to predict Day 2 and 5 embryo development to the blastocyst stage with an accuracy of 64%. Elevated concentrations of lysophosphatidylethanolamines (m/z = 453, 566, 588) in the culture media of Day 8 well-developing embryos were observed. Choline (104 m/z) and citrate (215 m/z) concentrations were increased in embryos in which development was retarded. Metabolic profiling provides possibilities to identify well-developing embryos before transfer, thus improving pregnancy rates and the number of calves born.
Additional keywords: developmental biology, embryology, IVF, IVM, reproduction.
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