Mizobe Yamato, Yoshida Mitsutoshi, Miyoshi Kazuchika
Laboratory of Animal Reproduction, Faculty of Agriculture, Kagoshima University, Kagoshima, Japan.
J Reprod Dev. 2010 Apr;56(2):285-90. doi: 10.1262/jrd.09-142a. Epub 2010 Jan 27.
The effects of mechanical vibration during in vitro maturation and/or in vitro culture after artificial activation of pig oocytes on maturation and development were examined. In addition, the optimal conditions were applied to in vitro production of blastocysts derived from miniature pig somatic cell nuclear transfer (SCNT) embryos. Mechanical vibration during in vitro maturation did not affect the rates (60.5 +/- 1.9-69.5 +/- 2.2%) of oocytes reaching the metaphase-II stage. However, the blastocyst formation rates after activation of oocytes matured with mechanical vibration for 5 sec at intervals of 30-60 min or for 10 sec at intervals of 60 min were significantly (P<0.05) higher than those of oocytes matured without mechanical vibration (25.7 +/- 2.0-28.1 +/- 2.7% vs. 12.3 +/- 1.4% and 25.8 +/- 1.8% vs. 15.7 +/- 1.9%, respectively). In contrast, mechanical vibration during in vitro culture after activation did not affect the blastocyst formation (11.6 +/- 5.2-16.5 +/- 3.0%) of oocytes. Mechanical vibration for 5 sec at intervals of 60 min during in vitro maturation of oocytes did not affect fusion (66.8 +/- 3.5-72.1 +/- 3.1%) with miniature pig somatic cells after enucleation. However, the blastocyst formation rate of SCNT embryos was improved (P<0.05) by mechanically vibrating recipient oocytes for 5 sec at intervals of 60 min during in vitro maturation, regardless of the presence or absence of the same treatment during in vitro culture (17.6 +/- 2.5% vs. 9.4 +/- 0.9% and 13.0 +/- 0.3% vs. 7.4 +/- 0.9%, respectively). The results indicated that mechanical vibration enhances the cytoplasmic maturation of in vitro-matured pig oocytes, resulting in improvement of their parthenogenetic development. In addition, it was shown that in vitro maturation of oocytes with mechanical vibration can be applied to efficient production of blastocysts derived from miniature pig SCNT embryos.
研究了猪卵母细胞人工激活后,体外成熟和/或体外培养过程中的机械振动对其成熟和发育的影响。此外,将最佳条件应用于源自小型猪体细胞核移植(SCNT)胚胎的囊胚的体外生产。体外成熟过程中的机械振动不影响卵母细胞达到减数分裂中期II期的比率(60.5±1.9 - 69.5±2.2%)。然而,以30 - 60分钟的间隔进行5秒机械振动成熟的卵母细胞,或60分钟间隔进行10秒机械振动成熟的卵母细胞,激活后的囊胚形成率显著(P<0.05)高于未进行机械振动成熟的卵母细胞(分别为25.7±2.0 - 28.1±2.7% 对12.3±1.4%以及25.8±1.8% 对15.7±1.9%)。相反,激活后体外培养过程中的机械振动不影响卵母细胞的囊胚形成(11.6±5.2 - 16.5±3.0%)。卵母细胞体外成熟过程中以60分钟间隔进行5秒机械振动不影响去核后与小型猪体细胞的融合率(66.8±3.5 - 72.1±3.1%)。然而,无论体外培养期间是否进行相同处理,体外成熟过程中以60分钟间隔对受体卵母细胞进行5秒机械振动可提高SCNT胚胎的囊胚形成率(P<0.05)(分别为17.6±2.5% 对9.4±0.9%以及13.0±0.3% 对7.4±0.9%)。结果表明,机械振动可增强体外成熟猪卵母细胞的细胞质成熟,从而改善其孤雌生殖发育。此外,研究表明,机械振动辅助的卵母细胞体外成熟可应用于高效生产源自小型猪SCNT胚胎的囊胚。