|dc.description.abstract||Selecting high quality preimplantation embryo for transfer has been the most difficult task when
producing embryos in vitro. To date the most used non-invasive method is based on visual
observation. Developing a non-invasive method for embryo assessment is essential to have a
profitable in vitro embryo production (IVP) and embryo transfer system. Molecular characterization
of embryo growth media has been proposed as an complementary method to visual assessment of
In this study we are demonstrating a novel method, allowing sample collection at different embryo
development stages, without compromising embryo quality, to determine potential viability markers
for bovine IVP.
Single bovine embryos were cultured in 60µl SOF+0.4% BSA droplets under mineral oil. Twenty µl of
culture media was removed at day 2, 5 and 8 post-fertilization. A total of 58 samples were analyzed
using liquid chromatography-mass spectrometry (Q-Trap 3200), followed by principal component
Our results indicate that there are significant differences (p<0,00001) in concentrations for proline
(m/z = 116), inositol (m/z of sodium adduct = 203) and citrate (m/z of sodium adduct = 215) also in
the amino acid group of leucine and isoleucine (m/z = 132), phenylalanine (m/z = 165) and arginine
(m/z = 211) between the normally developed and retarded in development embryo culture media.
Platelet activating factor (m/z = 524) (PAF) was roughly 3 fold increased in day 5 to day 8 embryo
culture media. Unfortunately the increase of PAF was not statistically significant between normally
developing and retarded embryos.
These results demonstrate that it is possible to remove culture media samples from droplets and not
significantly affect embryo development. Applying this method for embryo selection provides a
possibility to identify well-developing embryos and provides an opportunity for improving the herds