Abdi Shabnam, Salehnia Mojdeh, Hosseinkhani Saman
Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
Int J Fertil Steril. 2013 Oct;7(3):181-6. Epub 2013 Sep 18.
The aim of this study was to investigate follicular growth and steroid production in neonatal mouse ovary during in vitro culture.
In this experimental study, 7-day-old mouse whole ovaries were cultured in α-MEM (medium supplemented with 100 mIU/ml recombinant follicle stimulating hormone, 1% insulin, transferrin and selenium (ITS), 5% fetal bovine serum (FBS), 100 IU/ml penicillin and 50 μg/ml streptomycin for 7 days. The size of whole ovary was determined as mean area during culture. The survival rates of isolated preantral follicles after culture were assessed using trypan blue staining after being mechanically isolated. Histological evaluation of whole ovary was done by hematoxylin and eosin staining. 17-β estradiol, progesterone and dehydroepiandrosterone concentrations in the medium were measured during culture.
:The mean area of ovary increased after culture (1.47 vs. 0.21 mm(2)). The survival rate of isolated follicles in ovary after culture was 99.2%. There was a significant decline in the percentage of primordial follicles after seven days of culture (91.8 ± 0.2% vs. 65.1 ± 1.1%), whereas the rate of preantral follicles increased significantly (4.6 ± 0.4% vs. 29.2 ± 0.5%). The levels of estradiol, progesterone and dehydroepiandrosterone also increased significantly after culture (p<0.001).
These results show that the growth and development of primordial follicles in contrast with hormonal production decreased during in vitro culture of neonatal mouse ovaries.
本研究旨在调查新生小鼠卵巢在体外培养期间的卵泡生长和类固醇生成情况。
在本实验研究中，将7日龄小鼠的整个卵巢在α-MEM培养基（添加100 mIU/ml重组促卵泡激素、1%胰岛素-转铁蛋白-硒（ITS）、5%胎牛血清（FBS）、100 IU/ml青霉素和50 μg/ml链霉素）中培养7天。在培养期间将整个卵巢的大小确定为平均面积。在机械分离后，使用台盼蓝染色评估培养后分离的腔前卵泡的存活率。通过苏木精和伊红染色对整个卵巢进行组织学评估。在培养期间测量培养基中17-β雌二醇、孕酮和脱氢表雄酮的浓度。
培养后卵巢的平均面积增加（1.47对0.21平方毫米）。培养后卵巢中分离卵泡的存活率为99.2%。培养7天后原始卵泡的百分比显著下降（91.8±0.2%对65.1±1.1%），而腔前卵泡的比例显著增加（4.6±0.4%对29.2±0.5%）。培养后雌二醇、孕酮和脱氢表雄酮的水平也显著升高（p<0.001）。
这些结果表明，在新生小鼠卵巢的体外培养过程中，与激素生成相比，原始卵泡的生长和发育有所减少。
