Wongsrikeao Pimprapar, Otoi Takeshige, Taniguchi Masayasu, Karja Ni Wayan Kurniani, Agung Budiyanto, Nii Masaru, Nagai Takashi
Laboratory of Animal Reproduction, The United Graduate School of Veterinary Science, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.
Theriogenology. 2006 Jan 20;65(2):332-43. doi: 10.1016/j.theriogenology.2005.03.027. Epub 2005 Jun 20.
The objective was to determine the effects of supplementing hexoses in oocyte maturation and embryo culture medium on in vitro maturation (IVM) and in vitro fertilization (IVF) of porcine oocytes and in vitro development of in vitro produced (IVP) porcine embryos. In the first experiment, oocytes were matured in vitro in modified North Carolina State University (NCSU)-37 medium, supplemented with hexoses (glucose, fructose or galactose) at various concentrations: 0 (control), 2.5, 5.5 and 10 mM. Supplementing the maturation medium with either glucose or fructose (5.5 mM) increased the percentages of oocytes that matured to metaphase II (79.4 and 70.2%, respectively), as compared with the control group (P < 0.05). However, supplementing galactose had no effects on meiotic maturation and fertilization. In the second experiment, cleaved embryos were collected 3 days after IVF of oocytes matured in the maturation medium supplemented with 5.5 mM of glucose; they were cultured for an additional 4 days in modified NCSU-37 medium, supplemented with 5.5mM of glucose, fructose or galactose. The incidence of blastocyst formation was higher (P < 0.05) in the glucose and fructose groups (18.6 and 18.2%, respectively) than in the galactose group and non-supplemented control group (12.9 and 9.2%). Moreover, fructose supplementation increased the total cell number/blastocyst (48.0 versus 37.6) and reduced the index of DNA-fragmented nucleus in the blastocysts (7.6% versus 11.8%), as compared with glucose supplementation (P < 0.05). In conclusion, fructose was a practical alternative to glucose for supporting IVM of porcine oocytes and fructose was superior to glucose for producing high-quality porcine embryos in vitro.
本研究旨在确定在卵母细胞成熟和胚胎培养基中添加己糖对猪卵母细胞体外成熟（IVM）、体外受精（IVF）以及体外生产（IVP）猪胚胎体外发育的影响。在第一个实验中，卵母细胞在添加了不同浓度（0（对照组）、2.5、5.5和10 mM）己糖（葡萄糖、果糖或半乳糖）的改良北卡罗来纳州立大学（NCSU）-37培养基中进行体外成熟培养。与对照组相比，在成熟培养基中添加葡萄糖或果糖（5.5 mM）可提高成熟至中期II的卵母细胞百分比（分别为79.4%和70.2%）（P < 0.05）。然而，添加半乳糖对减数分裂成熟和受精没有影响。在第二个实验中，将在添加了5.5 mM葡萄糖的成熟培养基中成熟的卵母细胞进行体外受精，3天后收集分裂胚；将它们在添加了5.5 mM葡萄糖、果糖或半乳糖的改良NCSU-37培养基中再培养4天。葡萄糖组和果糖组的囊胚形成率较高（分别为18.6%和18.2%），高于半乳糖组和未添加组（分别为12.9%和9.2%）（P < 0.05）。此外，与添加葡萄糖相比，添加果糖可增加囊胚的总细胞数/囊胚（48.0对37.6）并降低囊胚中DNA片段化细胞核指数（7.6%对11.8%）（P < 0.05）。总之，果糖是支持猪卵母细胞体外成熟的葡萄糖的实用替代品，并且在体外生产高质量猪胚胎方面，果糖优于葡萄糖。
