Effect of cysteamine supplementation during in vitro culture of early stage bovine embryos on blastocyst rate and quality

Abstract

The aim of this study was to evaluate whether the addition of cysteamine to the in vitro culture media enhances the yield, hatching rate, total cell number and inner cell mass/total cell number ratio of bovine embryos. A total of 933 bovine oocytes collected from ovaries of 60 slaughtered donors were subjected to in vitro maturation and in vitro fertilization. Following fertilization, embryos were cultured in synthetic oviductal fluid without glucose. After 24 h embryos were transferred into synthetic oviductal fluid with 1.5 mM glucose and 0 (control), 50, 100 and 200 µM of cysteamine. After 48 h, the embryos were transferred into synthetic oviductal fluid with glucose but without cysteamine and cultured until Day 9. The number of cleaved embryos on Day 2, the total number of blastocysts on Day 7 and the number of hatched blastocysts on Day 9 were calculated. Differential staining of inner cell mass and trophectoderm cells of blastocysts were performed on Day 7 and Day 9 of in vitro culture. Supplementation of in vitro culture media with 100 µM cysteamine increased the blastocyst yield (P < 0.05) without affecting the hatching rate. Furthermore, the embryos cultured in the presence of 100 µM cysteamine had significantly higher number of inner cell mass cells (P < 0.05) and the proportion of inner cell mass cells (P < 0.05) compared with the controls. The results of the present study demonstrated that the addition of 100 µM cysteamine to the in vitro culture media improved blastocyst production rate and enhance embryo quality, which could lead to the improvement of the in vitro culture system for bovine embryos.

Antioxidant activity of oily extract obtained from Lippia origanoides improves the quality of bovine embryos produced in vitro

“…The blastocyst production rate in the culture medium containing 50μM/mL cysteamine, a standard antioxidant in commercial laboratory protocols used as a positive control in the present study, was 22.2±12.7%. This value is similar to the 29.1±2.7% of blastocysts in D7 found by Lojkic et al (2012) for zygotes grown at the same concentration of cysteamine. A blastocyst rate of 24% was obtained by Merton et al (2013) by adding 0.1mM cysteamine in oocyte maturation and in vitro culture media of bovine embryos.…”

In vitro production of early stage buffalo embryos in modified synthetic oviductal fluid (mSOF) medium

“…Higher cleavage percent obtained by earlier workers may be due to incorporation of various types of culture media as well as the supplements like βME, activin-A, EGF, IGF-1. Serrano et al (2012), Lojkic et al (2012) and Uger (2015) carried out the in vitro embryo culture in synthetic oviductal fluid medium as in present study but, Serrano et al (2012) added 30µl/ml βME to reduce the oxidative stress in presumptive zygotes during in vitro embryo culture. Lojkic et al (2012) supplemented 100 µM/ml cysteamine in the basic embryo culture media and obtained higher cleavage percent suggesting that early stage embryos are probably more sensitive to oxidative stress because of exhausted GSH pool synthesized during IVM.…”

“…Serrano et al (2012), Lojkic et al (2012) and Uger (2015) carried out the in vitro embryo culture in synthetic oviductal fluid medium as in present study but, Serrano et al (2012) added 30µl/ml βME to reduce the oxidative stress in presumptive zygotes during in vitro embryo culture. Lojkic et al (2012) supplemented 100 µM/ml cysteamine in the basic embryo culture media and obtained higher cleavage percent suggesting that early stage embryos are probably more sensitive to oxidative stress because of exhausted GSH pool synthesized during IVM. Uger (2015) added 10 µl/ml βME in the embryo culture media to reduce the oxidative stress.…”

Influence of Epidermal Growth Factor with Cysteamine on in -Vitro Buffalo Embryo Development

“…Addition of cysteamine enhanced the glutathione synthesis in maturation medium (De Matos and Furnus, 2000) to improve oocyte maturation by protecting oocytes from oxidative stress (Gasparrini et al, 2003). In the culture medium, cysteamine improved blastocyst production rate and enhanced embryo quality in various species as bovine (Lojkic et al, 2012) and buffalo (Ocampo and Ocampo, 2015). Moreover, it improved transformation efficiency of sperm head nucleus into male pronucleus during fertilization.…”