Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul 08826, Korea.
Center for Reproductive Sciences, Department of Obstetrics and Gynecology, University of California San Francisco, San Francisco, CA 94143, USA.
Int J Mol Sci. 2021 Jan 8;22(2):579. doi: 10.3390/ijms22020579.
An essential requirement for the success of in vitro maturation (IVM) of the oocyte is to provide an optimal microenvironment similar to in vivo conditions. Recently, somatic cell-based coculture or supplementation of a conditioned medium during IVM has been performed to obtain better quality of oocytes, because they mimic the in vivo reproductive tract by secreting paracrine factors. In this study, human adipose-derived stem cells (ASC) and their conditioned medium (ASC-CM) were applied to IVM of porcine oocytes to evaluate the effectiveness of ASC on oocyte development and subsequent embryo development. In results, both ASC and ASC-CM positively influence on oocyte maturation and embryo development by regulating growth factor receptors (, , and ), apoptosis (), cumulus expansion (, , and ), and oocyte maturation-related genes ( and ). In particular, the fluorescence intensity of GDF9 and BMP15 was markedly upregulated in the oocytes from the ASC-CM group. Furthermore, significantly high levels of growth factors/cytokine including VEGF, bFGF, IGF-1, IL-10, and EGF were observed in ASC-CM. Additionally, the ASC-CM showed active scavenging activity by reducing the ROS production in a culture medium. Consequently, for the first time, this study demonstrated the effect of human ASC-CM on porcine oocyte development and the alteration of mRNA transcript levels in cumulus-oocyte complexes.
卵母细胞体外成熟（IVM）成功的一个基本要求是提供类似于体内条件的最佳微环境。最近，已经进行了基于体细胞的共培养或在 IVM 期间补充条件培养基，以获得更好质量的卵母细胞，因为它们通过分泌旁分泌因子模拟体内生殖道。在这项研究中，将人脂肪来源干细胞（ASC）及其条件培养基（ASC-CM）应用于猪卵母细胞的 IVM，以评估 ASC 对卵母细胞发育和随后胚胎发育的有效性。结果表明，ASC 和 ASC-CM 通过调节生长因子受体（、、和）、凋亡（）、卵丘扩展（、、和）以及卵母细胞成熟相关基因（和），对卵母细胞成熟和胚胎发育产生积极影响。特别是，来自 ASC-CM 组的卵母细胞中 GDF9 和 BMP15 的荧光强度明显上调。此外，在 ASC-CM 中观察到包括 VEGF、bFGF、IGF-1、IL-10 和 EGF 在内的生长因子/细胞因子的水平显著升高。此外，ASC-CM 通过减少培养基中 ROS 的产生表现出积极的清除活性。因此，这项研究首次证明了人 ASC-CM 对猪卵母细胞发育的影响以及卵丘-卵母细胞复合物中 mRNA 转录水平的变化。
