Cui Xiang-Shun, Jin Yong-Xun, Shen Xing-Hui, Lee Jae Yeong, Lee Hyo-Sang, Yin Xi-Jun, Kong Il-Keun, Kim Nam-Hyung
Department of Animal Sciences, Chungbuk National University, Gaesin-dong, Cheongju, Chungbuk 361-763, South Korea.
Theriogenology. 2006 Jul 15;66(2):267-74. doi: 10.1016/j.theriogenology.2005.11.011. Epub 2006 Feb 20.
Despite many attempts to improve the in vitro maturation (IVM) of canine oocytes using various culture conditions, the efficiency of canine IVM remains very low compared with that of other domestic animals. In the present study we examined the effect of ovarian estrus stage on oocyte quality, and the effect of epidermal growth factor (EGF) in the presence and absence of macromolecules on the IVM of canine oocytes. More oocytes >or=100 microm in diameter were obtained from follicular ovaries than from ovaries at other estrus stages. After 72 h of culture, significantly more oocytes recovered from follicular ovaries than from anestrous and luteal ovaries were in germinal vesicle break down (GVBD). Bovine serum albumin (BSA) or fetal bovine serum (FBS) supplementation improved meiotic resumption as compared to polyvinyl alcohol (PVA) supplementation; however, there was no difference between the BSA and FBS supplements. The oocytes matured in North Carolina State University (NCSU) 37 medium containing 0.4% BSA and 100 ng/ml EGF showed the highest rates of development to the metaphase II (MII) stage when compared with the control treatment (P < 0.05). These results suggest that the estrous cycle of bitches influences the meiotic resumption of oocytes cultured in vitro, and EGF increases the meiotic resumption of canine oocytes in the presence of BSA in vitro.
尽管人们多次尝试通过各种培养条件来提高犬类卵母细胞的体外成熟(IVM)效率,但与其他家畜相比,犬类IVM的效率仍然很低。在本研究中,我们研究了卵巢发情阶段对卵母细胞质量的影响,以及在存在和不存在大分子的情况下表皮生长因子(EGF)对犬类卵母细胞IVM的影响。与处于其他发情阶段的卵巢相比,从卵泡期卵巢获得的直径≥100微米的卵母细胞更多。培养72小时后,从卵泡期卵巢回收的处于生发泡破裂(GVBD)状态的卵母细胞明显多于处于乏情期和黄体期卵巢的卵母细胞。与补充聚乙烯醇(PVA)相比,补充牛血清白蛋白(BSA)或胎牛血清(FBS)可改善减数分裂的恢复;然而,BSA和FBS补充剂之间没有差异。与对照处理相比,在含有0.4% BSA和100 ng/ml EGF的北卡罗来纳州立大学(NCSU)37培养基中成熟的卵母细胞发育到中期II(MII)期的比率最高(P < 0.05)。这些结果表明,母犬的发情周期会影响体外培养的卵母细胞的减数分裂恢复,并且在体外存在BSA的情况下,EGF可提高犬类卵母细胞的减数分裂恢复。