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利用生殖生物技术中重复使用的卵巢评估山羊腔前卵泡的体外培养系统:一种最大化繁殖潜力的替代方法。

Evaluation of in vitro culture systems for goat preantral follicles using reused ovaries from reproductive biotechniques: An alternative to maximize the potential of reproduction.

作者信息

Panta Amanda Michelle Torres, Silva Ana Flávia Bezerra da, Padilha Rodrigo Tenório, Correia Hudson Henrique Vieira, Rondina Davide, Figueiredo José Ricardo, Magalhães Padilha Deborah de Melo

机构信息

Department of Postgraduate in Biotechnology, Universidade Potiguar, Natal, Brazil.

Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), Faculdade de Veterinaria, State University of Ceará, Fortaleza, Brazil.

出版信息

Reprod Domest Anim. 2019 Mar;54(3):480-485. doi: 10.1111/rda.13377. Epub 2018 Dec 15.

Abstract

This study aimed to examine the in vitro culture of secondary preantral follicles, using reused ovaries, to compare both the 2D and 3D methods of in vitro culture of preantral follicles, and the system of medium replacement. Twenty-five pairs of ovaries from mixed-breed goats were used for the experiment. Follicular puncture of antral follicles was performed for in vitro production. After this procedure, the secondary preantral follicles were submitted to a microdissection procedure. The isolated preantral follicles were randomly divided into three treatments: (a) Two-dimensional culture with partial replacement of medium during culture (2D PR), (b) Three-dimensional culture with addition of medium during culture (3D AD) and (c) Three-dimensional culture with partial replacement of medium (3D PR). The culture period was 18 days. All treatments at the end of the in vitro culture period (18 days) presented a follicular survival rate which ranged from 59% to 70%, demonstrating that it was possible to perform an experiment with preantral follicles using ovaries that had previously been used in another reproductive biotechnique. The 3D AD treatment showed a survival percentage and follicular diameter higher than the 2D PR treatment, however, it did not differ from the 3D PR treatment. In conclusion, experiments employing the use of preantral follicles can be performed with success after the ovaries have been used for experiments with antral follicles. Moreover, the three-dimensional system with the addition of medium is recommended for in vitro culture of preantral follicles, since this system is more practical and financially feasible.

摘要

本研究旨在利用重复使用的卵巢对次级窦前卵泡进行体外培养,比较二维和三维窦前卵泡体外培养方法以及培养基更换系统。使用25对杂种山羊的卵巢进行实验。对窦卵泡进行卵泡穿刺以进行体外生产。在此过程之后,将次级窦前卵泡进行显微解剖操作。分离出的窦前卵泡随机分为三组处理:(a) 二维培养,培养期间部分更换培养基(二维部分更换,2D PR),(b) 三维培养,培养期间添加培养基(三维添加,3D AD),以及 (c) 三维培养,部分更换培养基(三维部分更换,3D PR)。培养期为18天。在体外培养期结束时(18天),所有处理组的卵泡存活率在59%至70%之间,这表明使用先前已用于另一生殖生物技术的卵巢对窦前卵泡进行实验是可行的。三维添加处理组的存活率和卵泡直径高于二维部分更换处理组,然而,与三维部分更换处理组没有差异。总之,在卵巢已用于窦卵泡实验后,使用窦前卵泡进行的实验能够成功进行。此外,推荐使用添加培养基的三维系统进行窦前卵泡的体外培养,因为该系统更实用且在经济上可行。

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