In vitro culture and non-invasive metabolic profiling of single bovine embryos

Reprod Fertil Dev. 2019 Jan;31(2):306-314. doi: 10.1071/RD17446.

Abstract

Selecting high-quality embryos for transfer has been a difficult task when producing bovine embryos invitro. 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, 20µL culture media was removed at Days 2, 5 and 8 after fertilisation from the same droplet (60µL). 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 (104m/z) and citrate (215m/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.

MeSH terms

  • Animals
  • Blastocyst / metabolism*
  • Cattle
  • Culture Media
  • Embryo Culture Techniques / veterinary*
  • Embryo Transfer / veterinary
  • Embryonic Development / physiology
  • Female
  • Mass Spectrometry
  • Metabolome*
  • Metabolomics
  • Pregnancy

Substances

  • Culture Media