Evaluación del KnockoutTM Serum Replacement en la producción de embriones, y expresión del gen Aquaporina 3 en blastocistos bovinos producidos in vitro post-congelación

Abstract

This research determined the effect of KnockoutTM Serum Replacement (KSR) on embryo production, and expression of the Aquaporin3 gene in bovine blastocysts produced in vitro post-freezing. The cumulus-oocyte complexes were matured and fertilized in vitro, the embryos were incubated for 6 days in culture medium supplemented with 3 treatments: 5% KSR, 5% fetal bovine serum (FBS), or 0.6% albumin bovine serum (CON). On day 7, after IVF, the embryos were classified according to their morphology. Quality 1 blastocysts were frozen and stored in liquid nitrogen. After being thawed, the total RNA was extracted, the complementary DNA was synthesized and the expression of the Aquaporin 3 gene was quantified by RT-qPCR. The data were processed using analysis of variance, the means compared with the least squares test. In the cleavage rate there was similarity between treatments, the proportion of total embryos ranged between: 24.9 to 26.3%; for blastocysts between 20.1 to 21.3%, and for early embryos: 31.1 to 40.4%. The KSR and SFB treatments obtained a higher rate of regular embryos than the CON; This difference was statistically significant (p<0.01). The CON group obtained the highest percentage of advanced embryos (45.9%) and the highest number of re-expanded blastocysts (7.6%) after thawing and incubation for 24 hours compared to the other treatments. Regarding the expression of Aquaporin3, there was no significant difference in the analysis between treatments