Effect of delipidant agents during in vitro culture on the development, lipid content, gene expression and cryotolerance of bovine embryos.

Abstract

In vitro produced embryos are still sensitive to the freezing process which can be explained, in part, by the high-lipid accumulation that characterizes these embryos. Therefore, we aimed to evaluate the effect of delipidating agents, L-carnitine and the trans-10 cis-12 conjugated linoleic acid (CLA) isomer, on blastocyst development, lipid content, gene expression and cryotolerance when added to embryo culture media. Embryos were cultured in four different media: T1: control (n=616), synthetic oviduct fluid (SOF) media with 5% foetal bovine serum (FBS); T2: L-carnitine (n=648), SOF medium with 5% FBS and 0.6mg/ml of L-carnitine; T3: CLA (n=627), SOF medium with 5% FBS and 100μM trans-10 cis-12 CLA; and T4: L-carnitine+CLA: (n=597), SOF medium with 5% FBS plus 0.6mg/ml L-carnitine and 100μM trans-10 cis-12 CLA. Supplementation of culture medium with either or both delipidating agents reduced (p<.05) blastocyst rate on D7 (T1=49±3.5; T2=39±3.0; T3=42±3.9 and T4=39±3.9), but did not affected gene expression (p>.05). Although embryos cultured in the presence of L-carnitine contained fewer (p<.05) lipid droplets than the control embryos, they showed a lower re-expansion rate 24hr post-thaw than those (p<.05). In conclusion, although L-carnitine reduced the amount of lipids in cultured embryos, the use of L-carnitine and CLA during in vitro culture was not able to improve the embryo production and the response to cryopreservation.