Porcine IVF embryo development and estrogen receptors are influenced by the concentration of percoll gradients during sperm selection

Abstract

This study aimed to evaluate the effect of three different concentrations of discontinuous gradients of percoll (90/45, 80/40, and 70/35) in the outcome of porcine in vitro fertilization (IVF) and its influence on further embryo development and quality. Embryo viability was assessed by the expression of estrogen receptors (E 2 R) and cleaved caspase-3 (CC3). The highest percoll concentration (90/45) resulted in the lowest embryo production (24.9%) in comparison with 80/40 (37.5%) and 70/35 (40.0%), with the production being similar between the two lowest concentrations. The hatching rate for 90/45 (26.2%) was lower than for 80/40 (45.5%), and both were similar to the Group 70/35 (32.9%). The hatched embryos from the concentration 90/45 showed the lowest proportion of E 2 R expression (3.6%), while the Groups 80/40 (22.6%) and 70/35 (39.3%) had a similar proportion of expression. The live embryos that did not hatch until Day 8 of culture presented a higher CC3 proportion for Group 90/45 (18.3%), in comparison with 80/40 (12.7%) and 70/35 (10.7%), with the latter two being similar. In conclusion, adjustments in percoll concentration used for sperm selection before porcine IVF can improve embryo production and competence for pregnancy recognition and establishment. K E Y W O R D S apoptosis, estrogen receptors, porcine IVF, sperm selection

Standardization of a Sex-Sorting Protocol for Stallion Spermatozoa by Means of Absolute RT-qPCR

“…Therefore, the above results indicate that the standard curves were correctly designed. swimming ability [5], and differential separation by percoll and albumin [6,7], among others. However, the methodology with the best accuracy currently (over 90%) is sexing by flow cytometry or sex-sorting, which is based on the difference in DNA content in Y chromosome with respect to the X chromosome [8].…”

“…Therefore, approaches to sexing offspring in mammals have focused on sexing male gametes, e.g., spermatozoa [3]. Methodologies that have been developed to achieve this goal include immunological detection techniques [4], differences in swimming ability [5], and differential separation by percoll and albumin [6,7], among others. However, the methodology with the best accuracy currently (over 90%) is sexing by flow cytometry or sex-sorting, which is based on the difference in DNA content in Y chromosome with respect to the X chromosome [8].…”

Osteopontin enhances sperm capacitation and in vitro fertilization efficiency in boars

“…The best approach is to use new technology to assist in reproduction. In vitro maturation (IVM) and in vitro fertilization (IVF) are techniques for the completion of fertilization in mammalian sperm and eggs in a manually controlled environment [ 4 , 5 ]. Although porcine embryos produced by IVM and IVF develop to the blastocyst stage, the low incidence of cleavage rate remains a significant impediment [ 6 ].…”