比较移植及体外培养对小鼠卵巢卵泡早期发育的影响-妇产科学专业论文.docxVIP

华中科技大学硕士学位论文 华 中 科 技 大 学 硕 士 学 位 论 文 PAGE 10 PAGE 10 第一部分 比较移植及体外培养对小鼠卵巢卵泡早期发育的影响 摘要 小鼠模型被广泛应用于卵巢始基卵泡激活（初始募集）基本规律及各种生长 分化因子调控作用的研究中。离开体内的自然环境，卵巢必须适应环境的改变， 比如二氧化碳浓度、营养供给、温度及酸碱度。更重要的是，离开体内调节系统 卵泡的生长会发生改变。近几年，卵巢的孵育方法主要包括体外培养和肾被膜下 移植。但没有文献报道不同孵育体系对卵巢发育的影响，及孵育卵巢和在体卵巢 间卵泡发育的区别。在本研究中，我们比较了体外培养卵巢、移植卵巢及体内生 长卵巢中各级卵泡的数量。用免疫组化技术和半定量逆转录聚合酶链反应 （RT-PCR）检测三个重要基因的表达及蛋白定位，包括透明体蛋白 3（ZP3）、生 长分化因子 9（GDF-9）和抗苗勒氏管激素（AMH）。结果显示，体外培养加速卵 泡激活、延缓发育卵泡的生长，影响 ZP3 、GDF-9 和 AMH 蛋白的表达模式。尽 管移植造成少量始基卵泡的丢失，但移植卵巢的特征与体内生长卵巢非常相似， 这一结果 提示移植 可以提供 最佳的卵 泡孵育环 境。在体 外培养体 系中， alpha-Modifie Eagle Medium （α-MEM）是较好的基础培养基。本实验系统研 究了孵育卵巢与体内生长卵巢间的相同及不同，证明了移植可以最好的模拟体内 卵巢生长环境。 关键词：培养体系、移植、卵巢、卵泡发育、自发激活 Characterization and comparison of early follicular growth in transplanted, in vitro cultured and in vivo grown mouse ovaries ABSTRACT Mouse model has been widely utilized to understand the basic principles of initial follicle recruitment (follicle activation) and the role of specific growth and differentiation factors. Outside of their natural environment, ovaries must cope with physical restriction. More importantly, eliminating in vivo regulation system may alter follicular growth. At present, the methods of ovarian incubation mainly include in vitro culture and transplantation under kidney capsule. But there were very few original research articles published on the comparison of different incubation systems and the differences between incubated and in vivo grown ovaries. In this study, we compared the numbers of follicles at different stages in cultured, transplanted and in vivo grown ovaries. Using immunohistochemical analysis and real-time polymerase chain reaction (PCR), we investigated three key genes expression and their protein location, including zona pellucida 3 (ZP3), growth and differentiation factor-9 (GDF-9) and anti-Müllerian Hormone (AMH). Our results suggest that in vitro culture accelerates follicle activation, retards the growth of developing follicles, and influences the expression patterns of ZP3, GD