Secondary follicle growth and oocyte maturation during encapsulated three-dimensional culture in rhesus monkeys: effects of gonadotrophins, oxygen and fetuin.
Secondary follicle growth and oocyte maturation during encapsulated three-dimensional culture in rhesus monkeys: effects of gonadotrophins, oxygen and fetuin.
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DOI：
10.1093/humrep/der049
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发表时间：
2011-05
影响因子：
6.1
通讯作者：
Jing Xu;M. Lawson;R. Yeoman;K. Pau;S. Barrett;M. Zelinski;R. Stouffer;R. Stouffer
中科院分区：
文献类型：
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作者：
Jing Xu;M. Lawson;R. Yeoman;K. Pau;S. Barrett;M. Zelinski;R. Stouffer;R. Stouffer
背景基于藻酸盐的基质支持非人类灵长类卵泡的三维（3D）结构，并且在FSH存在的情况下，允许窦前卵泡在体外发育至小窦阶段，包括产生卵巢类固醇和旁分泌因子。目前的研究调查了促性腺激素、胎球蛋白和氧气 (O2) 在体外改善灵长类卵泡生长和卵母细胞成熟的能力。方法从早期卵泡期卵巢中分离出猕猴次级卵泡，封装在海藻酸钠基质中，并在补充培养基中单独培养40天。检测了重组人（rh）FSH（高、中和低FSH分别为15、3和0.3ng/ml）、牛胎球蛋白（1或0mg/ml）和O2（5或20％v/v）的作用。每种培养条件下的一半卵泡在第 30-40 天接受 rhLH。达到窦期的卵泡用 rh 绒毛膜促性腺激素处理 34 小时以启动卵母细胞减数分裂成熟。分析培养基中的卵巢类固醇和抗苗勒氏管激素（AMH）。结果 培养条件的改善支持非人类灵长类动物的次级卵泡生长至窦期，并首次促进卵母细胞成熟至 MII 阶段。在 5% O2 存在胎球蛋白的情况下，如果用高或中 FSH 培养，卵泡的存活率最高，而在低 FSH 下，卵泡在第 5 周生长到更大的直径。 5% O2 下卵母细胞的健康和成熟得到促进。高 FSH 刺激卵泡生长产生类固醇，而 LH 则促进低 FSH 培养的卵泡的类固醇生成。与低 FSH 相比，高 FSH 下 AMH 生物合成升高，且在 5% O2 下比在 20% O2 下持续时间更长。结论 这种封装的 3D 培养模型允许进一步研究影响灵长类卵泡生长和卵母细胞成熟的内分泌和局部因素，与增强女性生育力保存选择相关。
BACKGROUND An alginate-based matrix supports the three-dimensional (3D) architecture of non-human primate follicles and, in the presence of FSH, permits the in vitro development of pre-antral follicles to the small antral stage, including the production of ovarian steroids and paracrine factors. The current study investigated the ability of gonadotrophins, fetuin and oxygen (O₂) to improve primate follicle growth and oocyte maturation in vitro. METHODS Macaque secondary follicles were isolated from the early follicular phase ovaries, encapsulated in a sodium alginate matrix and cultured individually for 40 days in supplemented medium. The effects of recombinant human (rh) FSH (15, 3 and 0.3 ng/ml for high, medium and low FSH, respectively), bovine fetuin (1 or 0 mg/ml) and O₂ (5 or 20% v/v) were examined. Half of the follicles in each culture condition received rhLH on Day 30-40. Follicles that reached antral stage were treated with rh chorionic gonadotrophin for 34 h to initiate oocyte meiotic maturation. Media were analyzed for ovarian steroids and anti-müllerian hormone (AMH). RESULTS Improved culture conditions supported non-human primate, secondary follicle growth to the antral stage and, for the first time, promoted oocyte maturation to the MII stage. In the presence of fetuin at 5% O₂, follicles had the highest survival rate if cultured with high or medium FSH, whereas follicles grew to larger diameters at Week 5 in low FSH. Oocyte health and maturation were promoted under 5% O₂. High FSH stimulated steroid production by growing follicles, and steroidogenesis by follicles cultured with low FSH was promoted by LH. AMH biosynthesis was elevated with high compared with low FSH and for longer under 5% O₂ than under 20% O₂. CONCLUSIONS This encapsulated 3D culture model permits further studies on the endocrine and local factors that influence primate follicle growth and oocyte maturation, with relevance to enhancing fertility preservation options in women.