title: Improvement of transgenic cloning efficiencies by culturing recipient oocytes and donor cells with antioxidant vitamins in cattle.
abstract: The present study was conducted to investigate effects of antioxidants during maturation culture of recipient oocytes and/or culture of gene-transfected donor cells on the meiotic competence of recipient oocytes, and the developmental competence and quality of the reconstructed embryos after nuclear transfer (NT) in cattle. Gene-transfected donor cells had negative effects on the proportions of blastocyst formation, total cell numbers, and DNA fragmentation indices of reconstructed embryos. Supplementation of either vitamin E (alpha-tocopherol: 100 microM) or vitamin C (ascorbic acid: 100 microM) during maturation culture significantly enhanced the cytoplasmic maturation of oocytes and subsequent development of embryos reconstructed with the oocytes and gene-transfected donor cells, but did not have synergistic effects. The supplementation of vitamin E during maturation culture of recipient oocytes increased the proportions of fusion and blastocyst formation of gene-transfected NT embryos, in which the proportions were similar to those of nontransfected NT embryos. When the gene-transfected donor cells that had been cultured with 0, 50, or 100 microM of vitamin E were transferred into recipient oocytes matured with vitamin E (100 microM), 50 microM of vitamin E increased the proportion of blastocyst formation and reduced the index of DNA fragmentation of blastocysts. In conclusion, gene-transfected donor cells have negatively influenced the NT outcome. Supplementation of vitamin E during both recipient oocyte maturation and donor cell culture enhanced the blastocyst formation and efficiently blocked DNA damage in transgenic NT embryos.
authors.affiliation: Laboratory of Animal Reproduction, The United Graduate School of Veterinary Science, Yamaguchi University, Yoshida, Yamaguchi, Japan.
authors.forename: Pimprapar
authors.initials: P
authors.lastname: Wongsrikeao
authors.forename: Takashi
authors.initials: T
authors.lastname: Nagai
authors.forename: Budiyanto
authors.initials: B
authors.lastname: Agung
authors.forename: Masayasu
authors.initials: M
authors.lastname: Taniguchi
authors.forename: Miho
authors.initials: M
authors.lastname: Kunishi
authors.forename: Shizuyo
authors.initials: S
authors.lastname: Suto
authors.forename: Takeshige
authors.initials: T
authors.lastname: Otoi
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D000975:Antioxidants / Q000494:pharmacology; D001205:Ascorbic Acid / Q000494:pharmacology*; D002417:Cattle / Q000235:genetics*; D002478:Cells, Cultured; D019976:Cloning, Organism / Q000379:methods*; D004338:Drug Combinations; D046149:Embryo Culture Techniques / Q000379:methods; D005260:Female; D005978:Glutathione / Q000032:analysis; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000737:chemistry / Q000187:drug effects*; D014801:Vitamin A / Q000494:pharmacology*
chemical_list: D000975:Antioxidants; D004338:Drug Combinations; D014801:Vitamin A; D005978:Glutathione; D001205:Ascorbic Acid