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离体山羊腔前卵泡的体外生长与成熟:胰岛素和促卵泡激素浓度、培养皿、共培养以及卵母细胞大小对减数分裂恢复的影响

In vitro growth and maturation of isolated caprine preantral follicles: Influence of insulin and FSH concentration, culture dish, coculture, and oocyte size on meiotic resumption.

作者信息

Silva G M, Brito I R, Sales A D, Aguiar F L N, Duarte A B G, Araújo V R, Vieira L A, Magalhães-Padilha D M, Lima L F, Alves B G, Silveira L B R, Lo Turco E G, Rodrigues A P, Campello C C, Wheeler M B, Figueiredo J R

机构信息

Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.

Postgraduate Biotechnology, Potiguar University/Laureate International Universities, Natal, CE, Brazil.

出版信息

Theriogenology. 2017 Mar 1;90:32-41. doi: 10.1016/j.theriogenology.2016.10.026. Epub 2016 Nov 9.

Abstract

The aims of this study were: (1) to evaluate the effect of different insulin concentrations, alone or in combination with either a fixed FSH concentration or increasing FSH concentrations on the in vitro culture of isolated caprine preantral follicles and (2) to analyze the efficiency of two IVM media and maturation culture systems (with or without coculture with in vivo grown oocytes) on the meiosis resumption. Secondary follicles were cultured for 18 days in a basic medium supplemented with low- or high-insulin concentration alone or with a fixed FSH concentration or with increasing FSH concentrations. Oocytes grown in vivo or in vitro were matured alone or cocultured. The high-insulin concentration associated with fixed FSH treatment had higher meiotic resumption rate (P < 0.05) and was the only treatment capable of producing oocytes in metaphase II. The rates of germinal vesicle, germinal vesicle breakdown, metaphase I, metaphase II (MII), meiotic resumption, and oocyte diameter were similar between the maturation media. In conclusion, a basic medium supplemented with 10-μg/mL insulin and 100-μg/mL FSH throughout the culture period improved meiotic resumption rate and produced MII oocytes from caprine preantral follicles cultured in vitro. The MII rate was similar between in vivo and in vitro grown oocytes ≥110 μm.

摘要

本研究的目的是

(1)评估不同胰岛素浓度单独使用或与固定促卵泡素(FSH)浓度或递增FSH浓度联合使用时,对分离的山羊腔前卵泡体外培养的影响;(2)分析两种体外成熟(IVM)培养基和成熟培养系统(有无与体内生长的卵母细胞共培养)对减数分裂恢复的效率。将次级卵泡在补充了单独的低或高胰岛素浓度、或固定FSH浓度、或递增FSH浓度的基础培养基中培养18天。体内或体外生长的卵母细胞单独成熟或共培养。与固定FSH处理联合使用的高胰岛素浓度具有更高的减数分裂恢复率(P < 0.05),并且是唯一能够产生处于中期II的卵母细胞的处理。成熟培养基之间的生发泡、生发泡破裂、中期I、中期II(MII)、减数分裂恢复和卵母细胞直径的比率相似。总之,在整个培养期补充10μg/mL胰岛素和100μg/mL FSH的基础培养基提高了减数分裂恢复率,并从体外培养的山羊腔前卵泡中产生了MII卵母细胞。体内和体外生长的≥110μm卵母细胞的MII率相似。

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