title: Glutathione and cysteine enhance porcine preimplantation embryo development in vitro after intracytoplasmic sperm injection.
abstract: 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.
authors.affiliation: Department of Animal Science and Biotechnology, College of Agriculture and Life Science, Chungnam National University, Daejeon, Korea.
authors.forename: Xiao Xia
authors.initials: XX
authors.lastname: Li
authors.affiliation: Institute of Agricultural Science, Chungnam National University, Daejeon, Korea.
authors.forename: Kyung-Bon
authors.initials: KB
authors.lastname: Lee
authors.affiliation: Department of Animal Science and Biotechnology, College of Agriculture and Life Science, Chungnam National University, Daejeon, Korea.
authors.forename: Ji Hye
authors.initials: JH
authors.lastname: Lee
authors.affiliation: Department of Animal Science and Biotechnology, College of Agriculture and Life Science, Chungnam National University, Daejeon, Korea.
authors.forename: Keun Jung
authors.initials: KJ
authors.lastname: Kim
authors.affiliation: Department of Animal Science and Biotechnology, College of Agriculture and Life Science, Chungnam National University, Daejeon, Korea.
authors.forename: Eun Young
authors.initials: EY
authors.lastname: Kim
authors.affiliation: Department of Animal Science and Biotechnology, College of Agriculture and Life Science, Chungnam National University, Daejeon, Korea.
authors.forename: Kil-Woo
authors.initials: KW
authors.lastname: Han
authors.affiliation: Department of Animal Science and Biotechnology, College of Agriculture and Life Science, Chungnam National University, Daejeon, Korea.
authors.forename: Kang-Sun
authors.initials: KS
authors.lastname: Park
authors.affiliation: Department of Animal Science and Biotechnology, College of Agriculture and Life Science, Chungnam National University, Daejeon, Korea.
authors.forename: Jung
authors.initials: J
authors.lastname: Yu
authors.affiliation: Department of Animal Science and Biotechnology, College of Agriculture and Life Science, Chungnam National University, Daejeon, Korea. Electronic address: kminkyu@cnu.ac.kr.
authors.forename: Min Kyu
authors.initials: MK
authors.lastname: Kim
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000187:drug effects*; D003470:Culture Media; D003545:Cysteine / Q000494:pharmacology*; D046149:Embryo Culture Techniques / Q000662:veterinary; D047108:Embryonic Development / Q000187:drug effects*; D005978:Glutathione / Q000494:pharmacology*; D017382:Reactive Oxygen Species; D020554:Sperm Injections, Intracytoplasmic / Q000379:methods / Q000662:veterinary*; D013552:Swine / Q000196:embryology*
chemical_list: D003470:Culture Media; D017382:Reactive Oxygen Species; D005978:Glutathione; D003545:Cysteine
keywords: Apoptosis; Cysteine; Glutathione (GSH); Intracytoplasmic sperm injection (ICSI); Reactive oxygen species (ROS)