Abdel-Ghani Mohammed A, Abe Yasuyuki, Asano Tomoyoshi, Hamano Seizo, Suzuki Hiroshi
Research Unit for Functional Genomics, National Research Center for Protozoan Diseases Obihiro University of Agriculture and Veterinary Medicine Inada-Choe 080-8555 Obihiro Japan.
Animal Bio-Technology Center Livestock Improvement Association 1-9-8, Kyobashi, Chuo-Ku Tokyo Japan.
Reprod Med Biol. 2010 Oct 26;10(1):43-49. doi: 10.1007/s12522-010-0069-y. eCollection 2011 Mar.
To investigate the ability of medium conditioned with bovine cumulus-oocyte complexes (COCs) to support nuclear maturation of canine oocytes recovered from domestic dog ovaries.
Cumulus-oocyte complexes were obtained from ovaries of domestic bitches (8 months old to 7 years old), and in-vitro maturation was evaluated in TCM-199 supplemented with different concentrations (0, 20, 30 or 50%) of bovine COCs-conditioned medium (BCM). The canine COCs were cultured for 72 or 96 h at 38.5°C in 5% CO, 5% O and 90% N. The bovine COCs-conditioned medium was obtained from culture of bovine COCs with TCM-199 supplemented with 5% FCS for 22 h at 38.5°C in 2% CO, 98% air.
The proportion of germinal vesicle breakdown (GVBD) after 72 h was significantly higher ( < 0.05) in medium supplemented with 30% BCM (20.7%) compared with the control group (13.4%). The rates of GVBD-MII stage were significantly higher ( < 0.05) when oocytes were matured with BCM at concentration of 30% (41.5%) compared with control (26.6%) after 72 h in-vitro culture. After 96 h in-vitro culture, the oocytes matured in medium supplemented with 30% BCM (5.5%) showed a significant increase ( < 0.05) in the proportion of MII compared with control (0.7%). However, increasing the cultivation time from 72 to 96 h resulted in an increase in oocyte degeneration rate.
The results suggested that bovine COCs-conditioned medium supplementation significantly increased nuclear maturation of canine oocytes.
研究用牛卵丘-卵母细胞复合体（COCs）处理过的培养基支持从家犬卵巢回收的犬卵母细胞核成熟的能力。
从8月龄至7岁的家犬卵巢中获取卵丘-卵母细胞复合体，并在添加不同浓度（0%、20%、30%或50%）牛COCs条件培养基（BCM）的TCM-199中评估体外成熟情况。将犬COCs在38.5°C、5%二氧化碳、5%氧气和90%氮气的环境中培养72或96小时。牛COCs条件培养基是通过将牛COCs与添加5%胎牛血清的TCM-199在38.5°C、2%二氧化碳、98%空气的环境中培养22小时获得的。
与对照组（13.4%）相比，添加30% BCM的培养基在72小时后生发泡破裂（GVBD）的比例显著更高（P<0.05）（20.7%）。在体外培养72小时后，与对照组（26.6%）相比，用30%浓度的BCM使卵母细胞成熟时，GVBD-MII期的比例显著更高（P<0.05）（41.5%）。在体外培养96小时后，与对照组（0.7%）相比，在添加30% BCM的培养基中成熟的卵母细胞MII期比例显著增加（P<0.05）（5.5%）。然而，将培养时间从72小时增加到96小时会导致卵母细胞退化率增加。
结果表明，添加牛COCs条件培养基可显著提高犬卵母细胞的核成熟率。
