Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceara, Fortaleza, CE, Brazil.
Cell Tissue Res. 2011 Nov;346(2):273-81. doi: 10.1007/s00441-011-1251-1. Epub 2011 Oct 11.
The aim of this study was to evaluate the effect of vascular endothelial growth factor-A(165) (VEGF-A(165)) on the in vitro development of goat secondary preantral follicles. Preantral follicles (≥150 μm in diameter) were isolated from the ovaries of adult mixed-breed goats and individually cultured for 18 days in αMEM in the absence (control) or presence of VEGF-A(165) at concentrations of 10 ng/ml (VEGF10) and 100 ng/ml (VEGF100). Analyses of follicular survival, diameter, antrum formation and rate of daily growth were performed every 6 days. At the end of the culture period, morphologically normal oocytes (≥110 μm in diameter) were taken for in vitro maturation (IVM). The results demonstrated that all follicles presented oocytes and granulosa cells that were morphologically normal and after labeling with calcein-AM, high rates of oocyte viability were observed in all treatments. The follicular diameter and the growth rate achieved in the presence of VEGF10 were higher than those of the control. Both treatments with VEGF-A(165) showed higher rates of oocyte recovery for IVM when compared with the control. Moreover, only the addition of VEGF-A(165) permitted oocytes grown in vitro to reach metaphase II. Thus, the addition of VEGF-A(165) to the culture medium improves the development of goat preantral follicles cultured in vitro, allowing the production of mature oocytes.
本研究旨在评估血管内皮生长因子 A(165)(VEGF-A(165))对山羊次级腔前卵泡体外发育的影响。腔前卵泡(直径≥150μm)从成年杂种羊卵巢中分离出来,在αMEM 中单独培养 18 天,在无(对照)或存在 VEGF-A(165)的情况下,浓度为 10ng/ml(VEGF10)和 100ng/ml(VEGF100)。每隔 6 天对卵泡存活率、直径、腔形成和日生长率进行分析。在培养期末,取形态正常的卵母细胞(≥110μm 直径)进行体外成熟(IVM)。结果表明,所有卵泡均有形态正常的卵母细胞和颗粒细胞,经 calcein-AM 标记后,所有处理的卵母细胞存活率均较高。在 VEGF10 存在的情况下,卵泡直径和生长速度均高于对照组。与对照组相比,两种 VEGF-A(165)处理均提高了 IVM 时的卵母细胞回收率。此外,只有添加 VEGF-A(165)才能使体外培养的卵母细胞达到中期 II 期。因此,在培养基中添加 VEGF-A(165)可改善体外培养的山羊腔前卵泡的发育,从而产生成熟的卵母细胞。