SummaryThis study aims to develop an in vitro co-culture system of in situ goat preantral follicles with bone marrow-derived mesenchymal stem cells (BM-MSC), evaluating the influence of these cells on follicular growth, rate of activation and morphologically normal follicles. Fragments of ovarian cortex were cultured for 1 or 7 days in the presence of BM-MSC (BM-MSC+) and absence of BM-MSC (BM-MSC−). Histological sections of the fragments were analysed and data were obtained regarding morphological classification, survival rate of morphologically normal follicles and rate of follicular activation. Culture medium on days 1 and 7 was also sampled for nitrite concentration and reduced glutathione activity. There was a reduction (P < 0.05) in the percentage of morphologically normal follicles in the BM-MSC+ compared with the fresh control only on the seventh day of culture. When comparing treatments, on the seventh day of culture, a higher rate of morphologically normal preantral follicles was observed in BM-MSC+ (P < 0.05). In both treatments, primordial and developing follicle rates were similar to the fresh control (P > 0.05). When comparing treatments with each other, as well as with the fresh control, no differences were observed in follicular diameter (P > 0.05) or nitrite concentration (P > 0.05). The concentration of reduced glutathione was lower on the seventh day of co-culture in both treatments (P < 0.05). In conclusion, co-culture had no influence on follicular or oocyte development. However, it was critical to maintain the survival of preantral follicles during 7 days of culture.

中文翻译：

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山羊窦前卵泡与骨髓间充质干细胞原位培养

摘要本研究旨在开发一种体外共培养体系原位山羊窦前卵泡与骨髓间充质干细胞 (BM-MSC)，评估这些细胞对卵泡生长、激活率和形态正常卵泡的影响。在存在 BM-MSC (BM-MSC+) 和不存在 BM-MSC (BM-MSC-) 的情况下，将卵巢皮质片段培养 1 或 7 天。分析碎片的组织切片并获得关于形态分类、形态正常卵泡的存活率和卵泡活化率的数据。还在第 1 天和第 7 天对培养基进行采样以检测亚硝酸盐浓度和降低的谷胱甘肽活性。有减少（磷< 0.05) BM-MSC+ 中形态正常卵泡的百分比与仅在培养第 7 天的新鲜对照相比。在比较处理时，在培养的第 7 天，在 BM-MSC+ 中观察到形态正常的窦前卵泡比例较高（磷< 0.05)。在这两种治疗中，原始和发育卵泡率与新鲜对照相似（磷> 0.05)。在比较治疗之间以及与新鲜对照时，卵泡直径没有观察到差异（磷> 0.05) 或亚硝酸盐浓度 (磷> 0.05)。两种处理共培养第 7 天还原型谷胱甘肽浓度均较低（磷< 0.05)。总之，共培养对卵泡或卵母细胞的发育没有影响。然而，在培养 7 天期间维持窦前卵泡的存活至关重要。