Arrivabene Neves Camila, Dos Santos Silva Lucilene, de Carvalho Camila Ernanda Sousa, Silva Carvalho Marina, Sarmento José Lindenberg Rocha, Vasconcelos Cavalcante Tânia, Arrivabene Mônica, Arrivabene Neves Tábatta, de Sousa Bezerra Maria Éllida, Júnior Antônio Luiz Gomes, da Silva Cleidson Manoel Gomes, de Carvalho Maria Acelina Martins
Federal University of Piauí, Integrated Nucleus of Morphology and Stem Cell Research (NUPCelt), Teresina, PI, Brazil.
Federal University of Piauí, Agricultural Science Centre, Teresina, PI, Brazil.
Zygote. 2020 Feb;28(1):65-71. doi: 10.1017/S0967199419000686. Epub 2019 Nov 18.
This 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.
本研究旨在建立一种原位山羊腔前卵泡与骨髓间充质干细胞（BM-MSC）的体外共培养系统，评估这些细胞对卵泡生长、激活率和形态正常卵泡的影响。卵巢皮质碎片在有BM-MSC（BM-MSC+）和无BM-MSC（BM-MSC-）的情况下培养1天或7天。对碎片的组织学切片进行分析，并获得有关形态学分类、形态正常卵泡的存活率和卵泡激活率的数据。还在第1天和第7天采集培养基样本，检测亚硝酸盐浓度和还原型谷胱甘肽活性。与新鲜对照组相比，仅在培养的第7天，BM-MSC+中形态正常卵泡的百分比有所降低（P<0.05）。在培养的第7天进行处理比较时，BM-MSC+中形态正常的腔前卵泡率更高（P<0.05）。在两种处理中，原始卵泡和发育中卵泡的比率与新鲜对照组相似（P>0.05）。在处理组之间以及与新鲜对照组进行比较时，卵泡直径（P>0.05）或亚硝酸盐浓度（P>0.05）均未观察到差异。在两种处理中，共培养第7天时还原型谷胱甘肽的浓度均较低（P<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7天的培养过程中维持腔前卵泡的存活至关重要。
