Kim Chung-Hoon, Cheon Yong-Pil, Lee You-Jeong, Lee Kyung-Hee, Kim Sung-Hoon, Chae Hee-Dong, Kang Byung-Moon
Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, College of Medicine, University of Ulsan, Asan Medical Center, Seoul 138-736, Republic of Korea.
Division of Development and Physiology, Department of Biology, Institute for Basic Science, Sungshin Women's University, Seoul 136-742, Republic of Korea.
Dev Reprod. 2013 Sep;17(3):269-74. doi: 10.12717/DR.2013.17.3.269.
This study was performed to evaluate the effects of fibroblast co-culture on in vitro maturation (IVM) of prepubertal mouse preantral follicles. The intact preantral follicles were obtained from the ovaries of 12-14 day old mice and these were cultured individually in α-minimal essential medium (α-MEM) supplemented with 5% fetal bovine serum (FBS), 100 mIU/Ml recombinant follicle stimulating hormone (rFSH), 1% insulin-transferrin-selenium, 100 μg/ml penicillin and 50 μg/Ml streptomycin as base medium for 12 days. A total of 200 follicles were cultured in base medium co-cultured with mouse embryonic fibroblast (MEF) (MEF group) (n=100) or only base medium as control group (n=100). Survival rate of follicles on day 12 of culture were significantly higher in the MEF group of 90.0%, compared with 77.0% of the control group (p=0.021). Follicle diameters on day 6 and 8 of the culture period were significantly larger in the MEF group than those in the control group (p=0.021, p=0.007, respectively). Estradiol levels in culture media on day 4, 6, 8, 10 and 12 of the culture period were significantly higher in the MEF group (p=0.043, p=0.021, p=0.006, p<0.001 and p=0.008, retrospectively). Our data suggest that MEF cell co-culture on IVM of mouse preantral follicle increases survival rate and promotes follicular growth and steroid production.
本研究旨在评估成纤维细胞共培养对青春期前小鼠窦前卵泡体外成熟（IVM）的影响。完整的窦前卵泡取自12 - 14日龄小鼠的卵巢，并将其单独培养于添加了5%胎牛血清（FBS）、100 mIU/Ml重组促卵泡激素（rFSH）、1%胰岛素 - 转铁蛋白 - 硒、100 μg/ml青霉素和50 μg/Ml链霉素的α - 最低必需培养基（α - MEM）中作为基础培养基，培养12天。总共200个卵泡被培养于与小鼠胚胎成纤维细胞（MEF）共培养的基础培养基中（MEF组）（n = 100），或仅培养于基础培养基中作为对照组（n = 100）。培养第12天卵泡的存活率在MEF组显著更高，为90.0%，而对照组为77.0%（p = 0.021）。培养期第6天和第8天的卵泡直径在MEF组显著大于对照组（分别为p = 0.021，p = 0.007）。培养期第4、6、8、10和12天培养基中的雌二醇水平在MEF组显著更高（回顾性分析，p = 0.043，p = 0.021，p = 0.006，p < 0.001和p = 0.008）。我们的数据表明，MEF细胞与小鼠窦前卵泡共培养可提高存活率并促进卵泡生长和类固醇生成。
