动态添加FSH对绵羊卵巢皮质组织体外培养的影响

The Effects of Dynamic Addition of FSH on Sheep Ovarian Cortical Tissue in vitro Culture

本研究旨在探索促卵泡素(FSH)对体外培养绵羊卵巢原始卵泡和组织活性的影响。通过稳定或动态方式添加不同浓度FSH(1、10、50、100ng·mL-1),用组织学和免疫组化PCNA方法来评估FSH对体外培养绵羊卵巢皮质组织中原始卵泡激活、生长、存活以及颗粒细胞增殖活性的影响;此外通过测定培养液中雌激素含量,评估卵巢组织体外培养过程中活性的保持。结果表明,先升后降动态添加FSH对于卵泡发育、存活和生长以及颗粒细胞的增殖具有全面的促进作用;同时组织分泌雌激素能力也高于其他组。以稳定方式添加时,50ng·mL-1FSH在培养1d时显示对卵泡发育的显著促进作用(P<0.05),添加10ng·mL-1以上浓度的FSH可以促进卵泡存活和生长(P<0.05)。在整个培养期内,各培养组中雌激素产量持续增加,说明卵巢活性在本培养体系中可以得到很好的维持。本试验结果表明FSH可能参与早期卵泡的发生,动态添加FSH更有利于体外培养卵泡的发育和组织活性的维持。

The objective of this work was to examine the effect of follicle-stimulating hormone（FSH） on the in vitro activation and viability of sheep primordial follicles.Sheep ovarian cortical fragments were cultured in a steel mesh culture system,various concentration of FSH（1,10,50,100 ng·mL-1） were either stabilized or dynamically（different days with different concentration of FSH） added in culture medium.Ovarian tissues,both fresh and cultured,were processed for histological and proliferating cell nuclear antigen（PCNA） assays,while tissue activity was evaluated by estrogen（E_2） production during culture duration.As results showed,FSH of dynamic addition FSH started with up-grade dose（1-100 ng·mL-1） followed by down-grade dose（100-1 ng·mL-1）,stimulated primordial follicle activation,survival,growth or granulosa cells proliferating,and also promoted the E_2 production in tissues in vitro cultured,compared to other groups.The concentration of FSH with 50 ng·mL-1 stimulated primordial follicle activation after 1 d culture,and in dose higher than 10 ng·mL-1 of FSH groups stimulated follicle survival and growth.During culture duration,E_2 production in all culture groups were increased constantly,which indicated that tissue activity were well maintained in the present study.In conclusion,this study suggestd that FSH involved in early follicle development and dynamic addition of FSH would benefit to primordial follicle developing and tissue activity maintenance.