In vitro maturation of canine oocytes co-cultured with bovine and canine granulosa cell monolayers
In vitro maturation of canine oocytes co-cultured with bovine and canine granulosa cell monolayers
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DOI：
10.1016/j.theriogenology.2011.08.007
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发表时间：
2012-01-15
期刊：
影响因子：
2.8
通讯作者：
Suzuki, Hiroshi
中科院分区：
文献类型：
--
作者：
Abdel-Ghani, Mohammed Ali;Shimizu, Takashi;Suzuki, Hiroshi
本研究探讨了牛颗粒细胞单层（BGML）和犬颗粒细胞单层（CGML）对犬卵母细胞核成熟的影响。将卵丘-卵母细胞复合体（COCs）或无卵丘的卵母细胞在Dulbecco改良Eagle培养基（DMEM，对照组）、含BGML的DMEM（BGML组）或含CGML的DMEM（CGML组）中在38.5 ℃、5%CO2下培养72小时。5%的氧气和90%的氮气所有培养基均补充有10%FCS、50 ng/mL EGF、2 μ g/mL雌二醇-17 β、0.1 IU/mL hCG、0.1 IU/mL FSH、0.25 mM甘草酸、100 μ M β-巯基乙醇、100 IU/mL青霉素和100 μ g/mL链霉素。在发情期和/或间情期从卵巢中回收的卵丘包裹卵母细胞中，BGML组的M-Ⅱ型卵母细胞比例最高（27.0%），CGML组为7.9%，对照组为3.5%（P < 0.05）。在从发情期和/或间情期卵巢采集的无卵丘卵母细胞中，与CGML共培养的M-H卵母细胞比例较低（3.0%），与对照组（3.0%）相似（P > 0.05）。但BGML的存在显著提高了裸卵发育为M-II的能力（P < 0.05）（10.2%）。BGML组减数分裂恢复率最高（P < 0.05），卵母细胞变性率最低（P < 0.05）。总之，BGML对犬卵母细胞的体外成熟系统以及减数分裂恢复具有积极的影响。(C)2012 Elsevier Inc. All rights reserved.
The present study investigated the effects of bovine granulosa cell monolayers (BGML) and canine granulosa cell monolayers (CGML) on nuclear maturation of canine oocytes with and without cumulus cells. Cumulus-oocyte complexes (COCs) or cumulus-free oocytes were cultured in Dulbecco's Modified Eagle's Medium (DMEM, control group), DMEM with BGML (BGML group), or DMEM with CGML (CGML group) for 72 h at 38.5 degrees C in 5% CO2. 5% O-2, and 90% N-2. All media were supplemented with 10% of FCS, 50 ng/mL of EGF, 2 mu g/mL of estradiol-17 beta, 0.1 IU/mL of hCG, 0.1 IU/mL of FSH, 0.25 mM of pyruvic acid, 100 mu M of beta-mercaptoethanol, 100 IU/mL of penicillin, and 100 mu g/mL of streptomycin. In cumulus-enclosed oocytes retrieved from ovaries at estrus and/or diestrus, the highest percentage of M-II oocytes (P < 0.05) was present in the BGML group (27.0%) compared with the CGML group (7.9%) and the control group (3.5%). In cumulus-free oocytes collected from ovaries at estrus and/or diestrus, the proportions of M-H oocytes co-cultured with the CGML were low (3.0%) and similar (P > 0.05) to proportions achieved with control (3.0%). However, the presence of BGML improved (P < 0.05) the ability of denuded oocytes to develop into M-II (10.2%). The BGML group had the highest overall meiotic resumption (P < 0.05), and least oocyte degeneration (P < 0.05) among experimental groups. In conclusion, BGML had a positive impact on the in vitro maturation system, as well as meiotic resumption of canine oocytes. (C) 2012 Elsevier Inc. All rights reserved.