Department of Obstetrics and Gynecology, Seoul National University Hospital, 101 Daehak-ro, Jongno-gu, Seoul, 03080, Korea.
Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, 103 Daehak-ro, Jongno-gu, Seoul, 03080, Korea.
Tissue Eng Regen Med. 2022 Feb;19(1):93-103. doi: 10.1007/s13770-021-00387-4. Epub 2021 Nov 6.
In vitro follicular maturation (IVFM) of ovarian follicles is an emerging option for fertility preservation. Many paracrine factors and two-dimensional or three-dimensional (3D) environments have been used for optimization. However, since most studies were conducted using the murine model, the physiological differences between mice and humans limit the interpretation and adaptation of the results. Marmoset monkey is a non-human primate (NHPs) with more similar reproductive physiology to humans. In this study, we attempted to establish a 3D matrix (Matrtigel)-based IVFM condition for marmoset ovarian follicles in combination with anti-apoptotic factor, X-linked inhibitor of apoptosis protein (XIAP).
Marmoset follicles were isolated as individual follicles and cultured in a single drop with the addition of 0, 10, and 100 μg/mL of XIAP molecules. Matured oocytes and granulosa cells from mature follicles were collected and analyzed. The average number of isolated follicles was less than 100, and primordial and antral follicles were abundant in the ovaries.
IVFM of marmoset follicles in 3D matrix conditions with XIAP increased the rates of survival and in vitro follicle development. Furthermore, oocytes from the 3D cultures were successfully fertilized and developed in vitro. The addition of XIAP increased the secretion of estradiol and aromatase. Furthermore, expression of granulosa-specific genes, such as bone morphogenetic protein 15, Oct4, and follicle-stimulating hormone receptor were upregulated in the in vitro-matured follicles than in normal, well-grown, and atretic follicles. Apoptosis-related B-cell lymphoma-2 was highly expressed in the atretic follicles than in the XIAP-treated follicles, and higher caspase-3 was localized in the XIAP-treated follicles.
In this study, we attempted to establish a 3D-matrix-based marmoset IVFM condition and demonstrated the synergistic effects of XIAP. The use of a 3D matrix may be applied as an optimal culture condition for marmoset ovarian follicles.
体外卵泡成熟（IVFM）是一种新兴的生育力保存选择。许多旁分泌因子和二维或三维（3D）环境已被用于优化。然而，由于大多数研究都是在小鼠模型中进行的，因此小鼠和人类之间的生理差异限制了对结果的解释和应用。狨猴是一种与人类生殖生理学更为相似的非人类灵长类动物（NHPs）。在这项研究中，我们试图建立一种基于 3D 基质（Matrigel）的狨猴卵巢卵泡 IVFM 条件，并结合抗凋亡因子 X 连锁凋亡抑制蛋白（XIAP）。
将狨猴卵泡作为单个卵泡分离出来，并在添加 0、10 和 100μg/ml XIAP 分子的单一液滴中进行培养。收集和分析成熟卵泡的成熟卵母细胞和颗粒细胞。分离的卵泡平均数量少于 100 个，卵巢中丰富的是原始卵泡和窦前卵泡。
在 3D 基质条件下添加 XIAP 可提高狨猴卵泡的体外存活率和卵泡发育率。此外，来自 3D 培养的卵母细胞在体外成功受精并发育。添加 XIAP 增加了雌二醇和芳香化酶的分泌。此外，与正常、良好生长和闭锁卵泡相比，体外成熟卵泡中颗粒细胞特异性基因骨形态发生蛋白 15、Oct4 和促卵泡激素受体的表达上调。与凋亡相关的 B 细胞淋巴瘤-2 在闭锁卵泡中的表达高于 XIAP 处理的卵泡，XIAP 处理的卵泡中定位的 caspase-3 更高。
在这项研究中，我们试图建立一种基于 3D 基质的狨猴 IVFM 条件，并证明了 XIAP 的协同作用。使用 3D 基质可能适用于狨猴卵巢卵泡的最佳培养条件。
