Jitjumnong Jakree, Tang Pin-Chi
Department of Animal Science, National Chung Hsing University, Taichung 40227, Taiwan.
The iEGG and Animal Biotechnology Center, National Chung Hsing University, Taichung 40227, Taiwan.
Animals (Basel). 2023 Mar 8;13(6):980. doi: 10.3390/ani13060980.
Multilayered secondary follicles were encapsulated in a 0.5% alginate matrix and cultured in a 3D culture system supplemented with bone morphogenetic protein 15 (BMP-15; 15 ng/mL) for 12 days. The in vitro development of ovarian follicles was evaluated. On day 12, the follicle diameter, follicle survival rate, and antrum formation rate were significantly higher for follicles cultured in BMP-15-supplemented medium than those cultured in regular medium. The percentage of ovulated metaphase II oocytes retrieved from follicles cultured in BMP-15-supplemented medium was greater than that of oocytes retrieved from follicles cultured in regular medium. The secretion of P4 was significantly higher on days 6, 8, and 10 in follicles cultured in BMP-15-supplemented medium. The result for E2 tended toward significance on day 12. Intracellular reactive oxygen species levels were higher and glutathione levels were lower in mature oocytes from the in vitro culture than in mature oocytes from an in vivo control. A 3D culture system using an alginate matrix and supplemented with BMP-15 effectively improves the outcomes of in vitro ovarian follicle culture.
多层次级卵泡被包裹在0.5%的藻酸盐基质中，并在添加了骨形态发生蛋白15（BMP-15；15 ng/mL）的三维培养系统中培养12天。评估了卵巢卵泡的体外发育情况。在第12天，添加BMP-15的培养基中培养的卵泡，其卵泡直径、卵泡存活率和窦腔形成率显著高于在常规培养基中培养的卵泡。从添加BMP-15的培养基中培养的卵泡中回收的排卵中期II期卵母细胞的百分比大于从常规培养基中培养的卵泡中回收的卵母细胞。在添加BMP-15的培养基中培养的卵泡，在第6、8和10天P4的分泌显著更高。在第12天，E2的结果有显著趋势。体外培养的成熟卵母细胞中细胞内活性氧水平较高，而谷胱甘肽水平低于体内对照的成熟卵母细胞。使用藻酸盐基质并添加BMP-15的三维培养系统有效地改善了体外卵巢卵泡培养的结果。
