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Effects of chlorogenic acid (CGA) supplementation during in vitro maturation culture on the development and quality of porcine embryos with electroporation treatment after in vitro fertilization
Authors:Thanh‐Van Nguyen  Manita Wittayarat  Lanh Thi Kim Do  Thanh Van Nguyen  Masahiro Nii  Zhao Namula  Toshiki Kunihara  Fuminori Tanihara  Maki Hirata  Takeshige Otoi
Affiliation:1. Faculty of Bioscience and Bioindustry, Tokushima University, Tokushima, Japan;2. Faculty of Veterinary Science, Prince of Songkla University, Songkhla, Thailand;3. Faculty of Veterinary Science, Vietnam National University of Agriculture, Ha Noi, Vietnam;4. Tokushima Prefectural Livestock Research Institute, Tokushima, Japan;5. College of Agricultural Science, Guangdong Ocean University, Guangdong, China
Abstract:Electroporation is the technique of choice to introduce an exogenous gene into embryos for transgenic animal production. Although this technique is practical and effective, embryonic damage caused by electroporation treatment remains a major problem. This study was conducted to evaluate the optimal culture system for electroporation‐treated porcine embryos by supplementation of chlorogenic acid (CGA), a potent antioxidant, during in vitro oocyte maturation. The oocytes were treated with various concentrations of CGA (0, 10, 50, and 100 μmol/L) through the duration of maturation for 44 hr. The treated oocytes were then fertilized, electroporated at 30 V/mm with five 1 msec unipolar pulses, and subsequently cultured in vitro until development into the blastocyst stage. Without electroporation, the treatment with 50 μmol/L CGA had useful effects on the maturation rate of oocytes, the total cell number, and the apoptotic nucleus indices of blastocysts. When the oocytes were electroporated after in vitro fertilization, the treatment with 50 μmol/L CGA supplementation significantly improved the rate of oocytes that developed into blastocysts and reduced the apoptotic nucleus indices (4.7% and 7.6, respectively) compared with those of the untreated group (1.4% and 13.0, respectively). These results suggested that supplementation with 50 μmol/L CGA during maturation improves porcine embryonic development and quality of electroporation‐treated embryos.
Keywords:antioxidant  chlorogenic acid  electroporation  embryo  oxidative stress
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