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来源于绵羊初级卵泡体外培养的 8 细胞孤雌胚胎。

Eight-cell parthenotes originated from in vitro grown sheep preantral follicles.

机构信息

Programa de Pós-Graduação em Ciências Veterinárias, Laboratório de Manipulação de Oócitos e Folículos Pré-Antrais, State University of Ceará, Fortaleza, Brazil.

出版信息

Reprod Sci. 2012 Nov;19(11):1219-25. doi: 10.1177/1933719112446072. Epub 2012 May 4.

Abstract

We investigated the effect of the leukemia inhibitory factor (LIF) alone or in association with follicle-stimulating hormone (FSH) on the in vitro growth and antrum formation of sheep preantral follicles. To evaluate oocyte quality, parthenogenetic activation of the oocytes recovered from in vitro grown preantral follicles was performed. Preantral follicles >110 μm in diameter were isolated and cultured for 18 days in basic medium either alone (control) or supplemented with LIF (10 or 50 ng/mL) in the absence or presence of FSH. Every 6 days the follicular survival, growth, and antrum formation were evaluated. When compared to control (P < .05), antrum formation was increased in follicles cultured in the presence of LIF10 and FSH. At the end of the culture, the oocytes underwent in vitro maturation (IVM); their viability and chromatin configuration were assessed. Although IVM was not affect by the treatments (P > .05), the numerically highest maturation rates (29.63%) were obtained when follicles were cultured in 50 ng/mL LIF (LIF50). Therefore, their oocytes were submitted to parthenogenetic activation; from which 58.3% of the mature oocytes resulted in 8-cell stage parthenotes. In conclusion, although LIF10 + FSH increases antrum formation when compared to a nonsupplemented medium (minimum essential medium), oocytes from sheep preantral follicles are capable of growing and maturing in vitro independent of LIF addition to the medium, which resulted in the formation of 8-cell parthenotes.

摘要

我们研究了白血病抑制因子 (LIF) 单独或与卵泡刺激素 (FSH) 联合对绵羊腔前卵泡体外生长和腔形成的影响。为了评估卵母细胞质量,对从体外生长的腔前卵泡中回收的卵母细胞进行了孤雌激活。分离直径 >110 μm 的腔前卵泡,并在基础培养基中培养 18 天,单独培养(对照)或添加 LIF(10 或 50 ng/mL),无论是否存在 FSH。每 6 天评估卵泡的存活、生长和腔形成。与对照组相比(P <.05),在添加 LIF10 和 FSH 的卵泡中培养时,腔形成增加。在培养结束时,卵母细胞进行体外成熟(IVM);评估其活力和染色质构型。尽管处理对 IVM 没有影响(P >.05),但当卵泡在 50 ng/mL LIF(LIF50)中培养时,获得了最高的成熟率(29.63%)。因此,将它们的卵母细胞进行孤雌激活;其中 58.3%的成熟卵母细胞形成 8 细胞期孤雌胚胎。总之,尽管 LIF10 + FSH 与未补充的培养基(最低必需培养基)相比增加了腔形成,但绵羊腔前卵泡的卵母细胞能够在不添加 LIF 的情况下进行体外生长和成熟,这导致了 8 细胞孤雌胚胎的形成。

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