Duarte Ana Beatriz Graça, Chaves Roberta Nogueira, Araújo Valdevane Rocha, Celestino Juliana Jales, Silva Gerlane Modesto, Lopes Cláudio Afonso Pinho, Tavares Líliam Mara Trevisan, Campelo Cláudio Cabral, de Figueiredo José Ricardo
Laboratory of Manipulation of Oocytes and Preantral Follicles, Veterinary Faculty, State University of Ceará, Fortaleza, Ceará, Brazil.
Zygote. 2011 Aug;19(3):215-27. doi: 10.1017/S0967199410000237. Epub 2010 Oct 28.
The aim of this study was to evaluate the influence of the number of follicles per drop (one or three) and antral follicles on in vitro development of isolated goat preantral follicles. Preantral follicles were isolated through microdissection and distributed individually (control) or in groups of three follicles (treatment) in microdroplets of α-MEM with or without 1000 ng/ml follicle stimulating hormone (FSH) for Experiments 1 and 2, respectively. Experiment 3 was divided into four treatments according to the presence of one or three preantral follicles, associated or not with antral follicles. After culture, oocytes were retrieved from morphologically normal follicles and submitted to in vitro maturation (IVM) and live/dead fluorescent labelling. Results of Experiment 1 (basic medium without FSH) showed that culture of preantral follicles in groups enhances viability, growth and antrum formation after 12 days. However, in the presence of FSH (Experiment 2), only the recovery rate of fully grown oocytes for IVM was significantly affected by grouping of follicles. In Experiment 3, in general, co-culture of preantral follicles with an early antral follicle had a detrimental effect on viability, antrum formation and production of oocytes for IVM. In conclusion, the performance of in vitro culture of goat preantral follicles is affected by the number of follicles per drop, the presence of an antral follicle and FSH.
本研究的目的是评估每滴卵泡数量（一个或三个）和窦卵泡对分离的山羊腔前卵泡体外发育的影响。通过显微解剖分离腔前卵泡，并分别（对照组）或三个卵泡一组（处理组）分布于含有或不含有1000 ng/ml促卵泡激素（FSH）的α-MEM微滴中，分别用于实验1和实验2。实验3根据一个或三个腔前卵泡的存在情况分为四种处理，有或没有窦卵泡。培养后，从形态正常的卵泡中取出卵母细胞，并进行体外成熟（IVM）和活/死荧光标记。实验1（不含FSH的基础培养基）的结果表明，腔前卵泡分组培养12天后可提高其活力、生长和腔形成。然而，在有FSH的情况下（实验2），只有IVM的完全成熟卵母细胞回收率受卵泡分组的显著影响。在实验3中，总体而言，腔前卵泡与早期窦卵泡共培养对活力、腔形成和IVM卵母细胞的产生有不利影响。总之，山羊腔前卵泡的体外培养性能受每滴卵泡数量、窦卵泡的存在和FSH的影响。
