Nagao Yoshikazu, Ohta Yuki, Murakami Hidemi, Kato Yoku
University Farm, Faculty of Agriculture, Utsunomiya University, Mohka, Tochigi, Japan.
Zygote. 2010 Nov;18(4):323-30. doi: 10.1017/S0967199409990396. Epub 2010 Mar 24.
In this study, we investigated the effects of methyl-β-cyclodextrin (MBCD) on in vitro fertilization and the subsequent development of bovine oocytes. Bovine oocytes matured in serum-free medium were inseminated with frozen-thawed sperm pre-incubated in protein-free modified Brackett and Oliphant medium (BO) containing various concentrations of MBCD for various periods. MBCD decreased the frequency of live sperm, however enhanced the capacitation and acrosome reaction of the live sperm. Pre-incubation of sperm with 0.5, 1.0 and 1.5 mM MBCD for 2 and 4 h increased the frequency of normal fertilization. Embryos derived from oocytes fertilized with spermatozoa pre-incubated with MBCD developed normally to the blastocyst stage and term. There were individual differences and similar tendencies in four different sires in terms of the effects of MBCD upon fertilization. These results indicate that the pre-incubation of bovine sperm with MBCD affects viability and capacitation status of the sperm and promotes fertilization in vitro. Embryos derived from oocytes fertilized with sperm pre-incubated with MBCD developed normally to the blastocyst stage and term.
在本研究中,我们调查了甲基-β-环糊精(MBCD)对牛卵母细胞体外受精及后续发育的影响。在无血清培养基中成熟的牛卵母细胞,用在含有不同浓度MBCD的无蛋白改良布拉克特和奥利芬特培养基(BO)中预孵育不同时间的冻融精子进行授精。MBCD降低了活精子的频率,但增强了活精子的获能和顶体反应。精子与0.5、1.0和1.5 mM MBCD预孵育2小时和4小时,增加了正常受精的频率。用与MBCD预孵育的精子使卵母细胞受精所产生的胚胎正常发育至囊胚期和足月。就MBCD对受精的影响而言,在四个不同种公牛中存在个体差异和相似趋势。这些结果表明,牛精子与MBCD预孵育会影响精子的活力和获能状态,并促进体外受精。用与MBCD预孵育的精子使卵母细胞受精所产生的胚胎正常发育至囊胚期和足月。