title: Temporal effects of alpha-tocopherol and L-ascorbic acid on in vitro fertilized porcine embryo development.
abstract: The susceptibility of embryos to reactive oxygen species (ROS) varies in different stages of embryo development. The present study evaluated temporal effects of alpha-tocopherol and L-ascorbic acid on the porcine embryo development, and investigated whether a single or twice supplements of these two antioxidants at a divided concentrations favors the embryo development. In order to determine temporal effects of alpha-tocopherol and/or L-ascorbic acid, 100 microM alpha-tocopherol or 200 microM L-ascorbic acid were supplemented to the North Carolina State University (NCSU)-23 embryo culture media at 0, 48, 96 and 120 h of culture. In another set of experiments, the concentration was divided into two equal halves, i.e., 50 microM alpha-tocopherol and 100 microM L-ascorbic acid, and supplemented twice at 0 and 48, 0 and 96, or 48 and 96 h of culture. Supplementing culture media with 100 microM alpha-tocopherol for the entire culture period of 168 h or starting from the 48 h of culture yielded higher blastocyst percentage compared with the control or starting from the 96 or 120 h of culture. L-Ascorbic acid (200 microM) alone or together with alpha-tocopherol (100 microM) with a single supplement did not affect the frequency of blastocyst formation or number of cells in blastocyst. L-ascorbic acid with a divided supplements yielded higher blastocyst percentage compared with the control. No synergistic effect was observed on embryo development at a single supplement of these antioxidants. Although, at divided supplements higher blastocyst percentage was observed compared with control group, no further beneficial effect was observed compared with alpha-tocopherol or L-ascorbic acid alone. Our results demonstrated that the embryotrophic effects of alpha-tocopherol and/or L-ascorbic acid, in terms of frequency of blastocyst formation and number of cells in blastocyst, depends on the concentration and supplementation timing.
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, 56-1 Sillim-dong, Gwanak-gu, Seoul 151-742, Republic of Korea.
authors.forename: Mohammad Shamim
authors.initials: MS
authors.lastname: Hossein
authors.forename: Md Abul
authors.initials: MA
authors.lastname: Hashem
authors.forename: Yeon Woo
authors.initials: YW
authors.lastname: Jeong
authors.forename: Myeong Seop
authors.initials: MS
authors.lastname: Lee
authors.forename: Sue
authors.initials: S
authors.lastname: Kim
authors.forename: Ji Hye
authors.initials: JH
authors.lastname: Kim
authors.forename: Ok Jae
authors.initials: OJ
authors.lastname: Koo
authors.forename: Seon Mi
authors.initials: SM
authors.lastname: Park
authors.forename: Eu Gine
authors.initials: EG
authors.lastname: Lee
authors.forename: Sun Woo
authors.initials: SW
authors.lastname: Park
authors.forename: Sung Keun
authors.initials: SK
authors.lastname: Kang
authors.forename: Byeong Chun
authors.initials: BC
authors.lastname: Lee
authors.forename: Woo Suk
authors.initials: WS
authors.lastname: Hwang
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000008:administration & dosage / Q000494:pharmacology; D001205:Ascorbic Acid / Q000008:administration & dosage / Q000494:pharmacology*; D004334:Drug Administration Schedule; D004622:Embryo, Mammalian / Q000187:drug effects* / Q000502:physiology; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D013552:Swine / Q000196:embryology*; D013997:Time Factors; D024502:alpha-Tocopherol / Q000008:administration & dosage / Q000494:pharmacology*
chemical_list: D000975:Antioxidants; D024502:alpha-Tocopherol; D001205:Ascorbic Acid