Laboratory of Manipulation of Oocytes and Preantral Follicles, Veterinary Faculty, State University of Ceara, Fortaleza, Brazil.
Cells Tissues Organs. 2010;192(4):272-82. doi: 10.1159/000317133. Epub 2010 Jun 26.
The aim of this study was to investigate the effects of nerve growth factor (NGF) on the in vitro culture of goat preantral follicles. Ovarian cortex fragments were cultured in α-MEM+ supplemented with 0, 1, 10, 50, 100 or 200 ng/ml NGF for 1 or 7 days. Small fragments of noncultured ovarian tissue as well as those cultured for 1 or 7 days were processed for histology and transmission electron microscopy. The results showed that after 1 or 7 days of culture at all concentrations of NGF, except at 1 ng/ml after 1 day of culture, there was a significant reduction in the percentage of normal follicles compared to noncultured tissues. At higher NGF concentrations (100 and 200 ng/ml) after 7 days of culture, there was a significant reduction in the percentage of normal follicles compared to tissues cultured in α-MEM+ alone or at the other concentrations of NGF. It is important to note that ultrastructural and fluorescent analyses confirmed only the integrity of follicles cultured with 1 ng/ml of NGF after 7 days. In contrast to noncultured control tissues, the percentage of developing follicles was significantly increased at all concentrations of NGF after 1 or 7 days of culture. We observed that follicular diameter was greater at 1 and 10 ng/ml NGF after culture for 7 days than at the other concentrations but was similar to follicles cultured in α-MEM+ alone. In conclusion, NGF improved the survival of goat preantral follicles cultured in vitro in a dose-dependent manner.
本研究旨在探讨神经生长因子(NGF)对山羊腔前卵泡体外培养的影响。将卵巢皮质碎片在添加 0、1、10、50、100 或 200ng/ml NGF 的α-MEM+中培养 1 或 7 天。未培养的卵巢组织以及培养 1 或 7 天的小组织片段均进行组织学和透射电子显微镜检查。结果表明,除培养 1 天的 1ng/ml 组外,在所有浓度的 NGF 培养 1 或 7 天后,正常卵泡的比例与未培养组织相比显著降低。在较高浓度的 NGF(100 和 200ng/ml)培养 7 天后,与单独在α-MEM+或其他浓度的 NGF 中培养的组织相比,正常卵泡的比例显著降低。值得注意的是,超微结构和荧光分析仅证实了在培养 7 天后用 1ng/ml NGF 培养的卵泡的完整性。与未培养的对照组织相比,在培养 1 或 7 天后,所有浓度的 NGF 均显著增加了发育中的卵泡的比例。我们观察到,在培养 7 天后,1 和 10ng/ml NGF 组的卵泡直径大于其他浓度组,但与单独在α-MEM+中培养的卵泡相似。总之,NGF 以剂量依赖的方式改善了山羊腔前卵泡的体外存活。