Abstract

Background: Insulin has been used as a stimulatory factor for in vitro cell culture since many years. Even for routine in vitro embryo production (IVP), insulin is added to the media during different steps. There is a strong difference in concentrations used in vitro compared to what is measured in vivo in follicular fluid or serum. We performed a pilot study on insulin stability to explain possible reasons for that variation. Results: We measured insulin concentrations before and after bovine oocyte maturation in an experiment by using a quantitative ELISA (Mercodia bovine insulin ELISA immunoassay) and found that concentrations were stable up to 22 h of incubation. We compared our results with eleven in vivo studies measuring insulin in either serum or follicular fluid and nine IVP-protocols using insulin. In all studies, in vitro concentrations were much higher compared with those found physiologically in vivo. Limited knowledge is available concerning the different activity and stability of insulin in vitro versus in vivo. Conclusions: The concentrations of insulin used in vitro are quite high in comparison to physiological concentrations found in serum or follicular fluid. One explanation may be a different stability or activity of insulin in vitro even if we could measure stable concentrations of insulin in our pilot study. More precise dose-effect studies have to be performed to draw clear conclusions about the consequences of the use of such high doses as they might have negative consequences for the developing embryo. Insulin has direct effects on the regulation of the metabolism and could even influence the epigenetic programming of the metabolism with unknown consequences for the offspring later in life.

Keywords

In vitro model; Blastocyst; Bovine; Embryo development; Oocyte maturation; Insulin

Published in

Acta Veterinaria Scandinavica
2016, Volume: 58, number: Supplement 1, article number: 66

SLU Authors

• Laskowski, Denise

  • Department of Clinical Sciences, Swedish University of Agricultural Sciences
    

• Sjunnesson, Ylva

  • Department of Clinical Sciences, Swedish University of Agricultural Sciences
    

• Gustafsson, Hans

  • Department of Clinical Sciences, Swedish University of Agricultural Sciences
    

• Humblot, Patrice

  • Department of Clinical Sciences, Swedish University of Agricultural Sciences
    

• Andersson, Göran

  • Department of Animal Biosciences, Swedish University of Agricultural Sciences
    

• Båge, Renee

  • Department of Clinical Sciences, Swedish University of Agricultural Sciences
    

UKÄ Subject classification

Genetics and Breeding


Publication identifier

DOI: https://doi.org/10.1186/s13028-016-0249-9

Permanent link to this page (URI)

https://res.slu.se/id/publ/77868