Mun Seong-Eun, Sim Bo-Woong, Yoon Seung-Bin, Jeong Pil-Soo, Yang Hae-Jun, Choi Seon-A, Park Young-Ho, Kim Young-Hyun, Kang Philyong, Jeong Kang-Jin, Lee Youngjeon, Jin Yeung Bae, Song Bong-Seok, Kim Ji-Su, Huh Jae-Won, Lee Sang-Rae, Choo Young-Kuk, Kim Sun-Uk, Chang Kyu-Tae
National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Chungcheongbuk-do, Republic of Korea.
Futuristic Animal Resource & Research Center, Korea Research Institute of Bioscience and Biotechnology, Chungcheongbuk-do, Republic of Korea.
PLoS One. 2017 Apr 13;12(4):e0175427. doi: 10.1371/journal.pone.0175427. eCollection 2017.
Despite the application of numerous supplements to improve in vitro culture (IVC) conditions of mammalian cells, studies regarding the effect of fetal bovine serum (FBS) on mammalian early embryogenesis, particularly in relation to redox homeostasis, are lacking. Herein, we demonstrated that early development of in vitro-produced (IVP) porcine embryos highly depends on the combination of FBS supplementation timing and embryonic reactive oxygen species (ROS) requirements. Interestingly, FBS significantly reduced intracellular ROS levels in parthenogenetically activated (PA) embryos regardless of the developmental stage. However, the beneficial effect of FBS on early embryogenesis was found only during the late phase (IVC 4-6 days) treatment group. In particular, developmental competence parameters, such as blastocyst formation rate, cellular survival, total cell number and trophectoderm proportion, were markedly increased by FBS supplementation during the late IVC phase. In addition, treatment with FBS elevated antioxidant transcript levels during the late IVC phase. In contrast, supplementation with FBS during the entire period (1-6 days) or during the early IVC phase (1-2 days) greatly impaired the developmental parameters. Consistent with the results from PA embryos, the developmental competence of in vitro fertilization (IVF) or somatic cell nuclear transfer (SCNT) embryos were markedly improved by treatment with FBS during the late IVC phase. Moreover, the embryonic stage-specific effects of FBS were reversed by the addition of an oxidant and were mimicked by treatment with an antioxidant. These findings may increase our understanding of redox-dependent early embryogenesis and contribute to the large-scale production of high-quality IVP embryos.
尽管应用了多种添加剂来改善哺乳动物细胞的体外培养(IVC)条件,但关于胎牛血清(FBS)对哺乳动物早期胚胎发育的影响,尤其是与氧化还原稳态相关的研究仍然匮乏。在此,我们证明了体外生产(IVP)的猪胚胎的早期发育高度依赖于FBS添加时间和胚胎活性氧(ROS)需求的组合。有趣的是,无论发育阶段如何,FBS都能显著降低孤雌激活(PA)胚胎中的细胞内ROS水平。然而,仅在后期(IVC 4 - 6天)处理组中发现FBS对早期胚胎发育具有有益作用。特别是,在IVC后期添加FBS可显著提高发育能力参数,如囊胚形成率、细胞存活率、总细胞数和滋养外胚层比例。此外,在IVC后期用FBS处理可提高抗氧化转录水平。相比之下,在整个时期(1 - 6天)或IVC早期(1 - 2天)添加FBS会极大地损害发育参数。与PA胚胎的结果一致,在IVC后期用FBS处理可显著提高体外受精(IVF)或体细胞核移植(SCNT)胚胎的发育能力。此外,添加氧化剂可逆转FBS对胚胎阶段的特异性影响,而用抗氧化剂处理可模拟这种影响。这些发现可能会增进我们对氧化还原依赖性早期胚胎发育的理解,并有助于大规模生产高质量的IVP胚胎。