Michel KERE(1), Chawalit SIRIBOON(1), Neng-Wen LO(2), Ngoc Tan NGUYEN(3) and Jyh-Cherng JU(1,4)
1) Department of Animal Science, National Chung Hsing University, Taichung 402, Taiwan, ROC
2) Department of Animal Science and Biotechnology, Tunghai University, Taichung 407, Taiwan, ROC
3) Agricultural Biotechnology Center, Institute of Agricultural Science for Southern Vietnam, Ho Chi Minh City, Vietnam
4) Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan, ROC
Abstract.
In this study, a dose-response assessment was performed to understand the relation between supplementation of media with L-ascorbic acid or vitamin C and porcine oocyte maturation and the in vitro development of parthenotes (PA) and handmade cloned (HMC) embryos. Various concentrations (0, 25, 50 and 100 µg/ml) of vitamin C supplemented in in vitro maturation (IVM) and culture (IVC) media were tested. None of these vitamin C additions affected nuclear maturation of oocytes, yet supplementation at 50 µg/ml led to significantly increased intracellular glutathione (GSH) levels and reduced reactive oxygen species (ROS). When cultured in IVM- and/or IVC-supplemented media, the group supplemented with 50 µg/ml of vitamin C showed improved cleavage rates, blastocyst rates and total cell numbers per blastocyst (P<0.05) compared with other groups (control, 25 µg/ml and 100 µg/ml). In contrast, supplementation with 50 µg/ml vitamin C decreased (P<0.05) the apoptosis index as compared with the groups supplemented with 100 µg/ml. In addition, even with a lower blastocyst rate to start with (37.6 vs. 50.3%, P<0.05), supplementation of HMC embryos with vitamin C meliorated their blastocyst quality to the extent of PA embryos as indicated by their total cell numbers (61.2 vs. 59.1). Taken together, an optimized concentration of vitamin C supplementation in the medium not only improves blastocyst rates and total cell numbers but also reduces apoptotic indices, whereas overdosages compromise various aspects of the development of parthenotes and cloned porcine embryos.
Key words: Culture system, Embryo development, Handmade cloning, Parthenote, Vitamin C
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