Glutathione and cysteine enhance porcine preimplantation embryo development in vitro after intracytoplasmic sperm injection

Because intracytoplasmic sperm injection (ICSI) had been introduced to animal science, not only reproductive biology of domestic animals, but also medicine to treat infertility has been developed. This assisted reproductive technology is beneficial for generating transgenic animals, especially pigs,...

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Published in:Theriogenology 2014-01, Vol.81 (2), p.309-314
Main Authors: Li, Xiao Xia, Lee, Kyung-Bon, Lee, Ji Hye, Kim, Keun Jung, Kim, Eun Young, Han, Kil-Woo, Park, Kang-Sun, Yu, Jung, Kim, Min Kyu
Format: Article
Language:English
Subjects:
Animals
Apoptosis
Blastocyst - drug effects
Culture Media
Cysteine
Cysteine - pharmacology
Embryo Culture Techniques - veterinary
Embryonic Development - drug effects
Glutathione (GSH)
Glutathione - pharmacology
Intracytoplasmic sperm injection (ICSI)
Reactive Oxygen Species
Reactive oxygen species (ROS)
Sperm Injections, Intracytoplasmic - methods
Sperm Injections, Intracytoplasmic - veterinary
Swine - embryology
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Summary:Because intracytoplasmic sperm injection (ICSI) had been introduced to animal science, not only reproductive biology of domestic animals, but also medicine to treat infertility has been developed. This assisted reproductive technology is beneficial for generating transgenic animals, especially pigs, because polyspermy is the greatest hurdle in porcine IVF when researchers make highly qualified preimplantation embryos. However, ICSI-derived embryos expressed high level of reactive oxygen species (ROS), which are known to cause serious dysfunction during preimplantation development. The objective of this study was to investigate the developmental competence, ROS level, and apoptosis index when glutathione (GSH) or cysteine was supplemented into the in vitro culture medium for ICSI-derived porcine embryos. First, we evaluated the effect of different concentrations of GSH or cysteine on developmental ability of porcine ICSI-derived embryos. The cleavage rate (79.6%) and the blastocyst formation rate (20.9%) were significantly improved in culture medium supplemented with 1 mmol/L GSH compared with other concentrations or no supplementation. Also, 1.71 mmol/L cysteine showed a significantly higher proportion of cleavage (80.7%) and blastocyst formation (22.5%) than other cysteine-supplemented groups. Next, we confirmed that intracellular ROS level was significantly reduced in the group of blastocysts cultured with GSH or cysteine after ICSI compared with the no supplementation group. Finally, we found that terminal uridine nick-end labeling index, fragmentation, and total apoptosis were significantly decreased and the total cell number was significantly increased in blastocysts when ICSI-derived embryos were cultured with supplementation of 1.71 mmol/L cysteine or 1 mmol/L GSH. Taken together, these results strongly indicate that GSH or cysteine can improve the developmental competence of porcine ICSI-derived embryos by reducing intracellular ROS level and the apoptosis index.
ISSN:0093-691X
1879-3231
DOI:10.1016/j.theriogenology.2013.09.030