1Department of Obstetrics and Gynecology, Seoul National University College of Medicine, 101 Daehak-ro, Jongno-Gu, Seoul, 03080 Republic of Korea.
2Department of Obstetrics and Gynecology, Korea University Guro Hospital, 148 Gurodong-ro, Guro-gu, Seoul, 08308 Republic of Korea.
Tissue Eng Regen Med. 2019 May 11;16(3):275-283. doi: 10.1007/s13770-019-00188-w. eCollection 2019 Jun.
Vitamin is a well-known co-factor for many metabolic processes and its roles in fertility and follicular growth have been studied. Vitamin supplementation is frequently achieved by daily ingestion in the form of a complex capsule. However, the role of single and complex vitamins in maturation of murine follicles is not fully elucidated.
In this study, we evaluated the effects of two forms of vitamins. Pure L-ascorbic acid, and multi-vitamin (vitamin C + vitamin B complex) was treated at two different concentrations (50 and 100 µg/ml), to pre-puberty murine follicles during maturation. To determine the specific stage of growth that is affected by treatment with vitamins, the vitamins were treated from day 0, 4, 9, and 13. Growth of each follicle was assessed by measuring diameters of whole expanded area and of the granulosa cells. Expression of follicular and oocyte growth-related genes and the effect of vitamin on the viability of follicles was assessed using senescence associated β-galactosidase staining.
Treatment with vitamins promoted the growth of murine follicles and the upregulated the expression of granulosa cell- and oocyte-specific genes such as , , and . The proliferation of the granulosa cells was enhanced by the treatment of vitamin. Fifty µg/ml concentration vitamin showed greater effects compared to higher concentration. The viability of grown follicles was also significantly improved in vitamin-treated follicles. The effects of single L-ascorbic acid and complex vitamin were not significantly different to those of day 4 and day 9 follicles. Vitamins promoted murine follicle development with different effects on specific growth stage.
Supplementation of vitamins during maturation of murine follicles is an efficient strategy for expansion of follicular cells. These results could be customized to the sophisticated culture of follicles retrieved from aged or cancer-survived female that contain smaller number of follicles with reduced potential to develop into mature follicles.
维生素是许多代谢过程的已知辅助因子，其在生育和卵泡生长中的作用已得到研究。维生素补充剂通常以复合胶囊的形式每天摄入来实现。然而，单一和复合维生素在小鼠卵泡成熟中的作用尚未完全阐明。
在这项研究中，我们评估了两种形式的维生素的作用。纯 L-抗坏血酸和多种维生素（维生素 C + 维生素 B 复合物）以两种不同浓度（50 和 100 µg/ml）处理青春期前的小鼠卵泡在成熟过程中。为了确定受维生素处理影响的特定生长阶段，维生素从第 0、4、9 和 13 天开始处理。通过测量整个扩张区域和颗粒细胞的直径来评估每个卵泡的生长。使用衰老相关的β-半乳糖苷酶染色评估卵泡和卵母细胞生长相关基因的表达以及维生素对卵泡活力的影响。
维生素处理促进了小鼠卵泡的生长，并上调了颗粒细胞和卵母细胞特异性基因的表达，如 、 和 。维生素处理增强了颗粒细胞的增殖。50 µg/ml 浓度的维生素比高浓度的维生素表现出更大的效果。维生素处理的卵泡中生长卵泡的活力也显著提高。与第 4 天和第 9 天的卵泡相比，单一的 L-抗坏血酸和复合维生素的作用没有显著差异。维生素以不同的方式对特定的生长阶段发挥作用，促进了小鼠卵泡的发育。
在小鼠卵泡成熟过程中补充维生素是扩大卵泡细胞的有效策略。这些结果可以针对从老年或癌症幸存女性中获得的卵泡的复杂培养进行定制，这些卵泡中卵泡数量较少，发育成成熟卵泡的潜力降低。
