Abdi Shabnam, Salehnia Mojdeh, Hosseinkhani Saman
Anatomy Department Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Anatomy Department Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Reprod Biomed Online. 2015 May;30(5):493-503. doi: 10.1016/j.rbmo.2015.01.009. Epub 2015 Jan 30.
To investigate the development of follicles and incidence of apoptosis in vitrified cultured mouse ovaries in the presence and absence of Kit ligand, 1-week-old mouse ovaries were cultured in the presence or absence of Kit ligand for 7 days. Development and function of ovarian follicles was evaluated by histology and hormonal analysis. Apoptosis assessment was conducted by analysis of DNA laddering, TdT (terminal deoxynucleotidyl transferase)-mediated dUDP nick-end-labelling and caspase-3/7 activity. The proportion of preantral follicles and the level of 17-β oestradiol, progesterone and dehydroepiandrosterone were increased in all cultured groups, and it was significantly higher in Kit ligand treated groups than in the control (P < 0.001). The number of apoptotic signals in both vitrified samples is significantly higher than in the non-vitrified control (P < 0.01), and these signals are significantly lower in both Kit ligand treated groups than in non-Kit ligand treated groups (P < 0.001). The level of caspase-3/7 activity was higher in vitrified cultured ovaries than non-vitrified group (P < 0.01). Kit ligand was shown to improve in-vitro development of follicles, and also acted as an anti-apoptotic factor in vitrified ovaries. The developmental potential of follicles in vitrified groups was lower than that in fresh ovaries.
为了研究在有或没有Kit配体的情况下玻璃化培养的小鼠卵巢中卵泡的发育情况和细胞凋亡发生率,将1周龄小鼠的卵巢在有或没有Kit配体的情况下培养7天。通过组织学和激素分析评估卵巢卵泡的发育和功能。通过分析DNA梯状条带、TdT(末端脱氧核苷酸转移酶)介导的dUDP缺口末端标记和caspase-3/7活性来进行细胞凋亡评估。所有培养组中窦前卵泡的比例以及17-β雌二醇、孕酮和脱氢表雄酮的水平均有所增加,且Kit配体处理组显著高于对照组(P<0.001)。两个玻璃化样本中的凋亡信号数量均显著高于未玻璃化对照组(P<0.01),且两个Kit配体处理组中的这些信号均显著低于未用Kit配体处理的组(P<0.001)。玻璃化培养的卵巢中caspase-3/7活性水平高于未玻璃化组(P<0.01)。结果表明,Kit配体可改善卵泡的体外发育,并且在玻璃化卵巢中起到抗凋亡因子的作用。玻璃化组中卵泡的发育潜能低于新鲜卵巢中的卵泡。