Effect of capacitating agents on sperm pretreatment during in vitrofertilization for blastocyst production in caprines

Abstract

The present study aimed at comparing the relative efficacy of different capacitating agents used for sperm pretreatment during in vitro fertilization (IVF) and subsequent embryo development of caprine oocytes. In experiment 1, the effect of different concentrations of heparin (0, 20, and 50 µg/mL) on sperm pretreatment for different periods of time (30 and 60 min) was studied. In experiment 2, the dose-dependent effect of calcium ionophore (0, 0.1, 0.2, and 0.5 µM) on sperm pretreatment was assessed. Experiment 3 validated and compared the effects of both capacitating agents on sperm pretreatment for capacitation based on the results of the first two experiments. The treated sperms from each group were used for in vitro fertilization and subsequent embryo development. The results indicated that in experiment 1, capacitation with heparin at 20 µg/mL for 60 min had significantly higher (P < 0.05) cleavage and blastocyst production. In experiment 2, capacitation with calcium ionophore at 0.1 µM had significantly more morulae (P < 0.05) and numerically more blastocyst production. In experiment 3, capacitation with heparin at 20 µg/mL had a significantly higher (P < 0.05) blastocyst production rate as compared to calcium ionophore treatment. Based on this study, heparin can be used to enhance capacitation of freshly collected sperms during in vitro fertilization and subsequent embryo development.

Effect of Foetal Bovine Serum on sperm motility, acrosome reaction and spermatic interaction to zona pellucida in alpacas (Vicugna pacos)

“…The use of foetal bovine serum in the incubation medium of sperm has not been common; for this reason, little is known about the effect of FBS on sperm functionality. However, some studies have used FBS as a supplement in sperm incubation and in vitro fertilization (Chen et al., ; Funahashi & Day, ,b; Goel et al., ; González‐Fernández et al., ; Karimi et al., ; Keskintepe et al., ; Leibfried‐Rutledge et al., ; Tian et al., ; Yang et al., ).…”

“…In the present work, the effect of 10% FBS on the functionality of the alpaca sperm was determined. This concentration has been previously used in the sperm incubation medium of other species (Chen et al., ; Funahashi & Day, ,b; Goel et al., ; González‐Fernández et al., ; Kwon et al., ; Yang et al., ). Rizal and Sangadji () observed that the concentration of FBS at 8%, 10% and 12% did not present significant differences in relation to the sperm motility of ram.…”

“…This suggests that FBS may initiate some mechanisms linked to sperm capacitation in alpaca. However, in previous studies, where the FBS was supplemented in the incubation medium, the sperm capacitation was induced by synthetic substances such as calcium ionophore (Chen et al., ; Goel et al., ; Keskintepe et al., ; Long et al., ) or heparin (Goel et al., ; Kwon et al., ), and the FBS was not considered as an inductor of the sperm capacitation, although it may have had an adjuvant effect in this process. In the work performed by Long et al.…”

“…In the work done by Goel et al. (), heparin or calcium ionophore was used to capacitate goat sperm and which were then incubated with oocytes in a medium with 10% FBS, resulting in embryos. On the other hand, in this same work, cleavage did not occur when sperm were only incubated with FBS.…”

“…The use of foetal bovine serum (FBS) in reproductive technologies such as embryo culture and oocyte maturation has been extensively used (Goel, Goel, Bhatiya, & Kharche, 2016;González-Fernández, Macedo, Lopes, Rocha, & Macías-García, 2015;Leibfried-Rutledge, Critser, & First, 1986;Rehman, Collins, Suh, & Wright, 1994;Wang, Liu, Holyoak, & Bunch, 1997;Yang, Chen, & Li, 1999). However, supplementation of FBS in the incubation medium for sperm has been less frequent.…”