School of Life Science and Biotechnology, College of Natural Sciences, Kyungpook National University, 1370 Sankyuk-dong, Buk-ku, Daegu, 702-701, Republic of Korea.
In Vitro Cell Dev Biol Anim. 2010 Feb;46(2):148-54. doi: 10.1007/s11626-009-9250-1.
The major objective of this study was to improve the development rate of parthenogenetic porcine embryos. In this study, the anti-oxidative and anti-apoptotic effects of three antioxidants, β-mercaptoethanol (β-ME), α-tocopherol, and extracellular superoxide dismutase (EC-SOD), were examined on the development of parthenogenetic porcine embryos. The development rate of parthenogenetic porcine embryos to the blastocyst stage was 8.1% for control; 19.1%, 14.6%, and 5.0% for 1, 3, and 5 μM β-ME; 17.2% and 17.5% for 50 and 100 μM α-tocopherol and 12.0% and 4.0% for EC-SOD transgenic mouse embryonic fibroblast (Tg-MEF) and EC-SOD non-transgenic mouse embryonic fibroblast (NTg-MEF) conditioned medium at day 3, respectively. Here, β-ME, α-tocopherol, and EC-SOD Tg-MEF conditioned medium increased the development rate of parthenogenetic porcine embryos to the blastocyst stage (P < 0.05). The average number of total cells and apoptotic cells at the blastocyst was analyzed at the optimal conditions of the three antioxidants. The three antioxidants increased the average number of total cells at the blastocyst, and they decreased apoptotic cells at the blastocyst as compared to control without supplementation (P < 0.05). When the reactive oxygen species levels in two-cell embryos after 1 μM β-ME and 100 μM α-tocopherol treatment were examined, those were lower than control group (P < 0.05). In conclusion, it was found that the three antioxidants, β-mercaptoethanol, α-tocopherol, and EC-SOD Tg-MEF, conditioned medium can play a role as a strong stimulator in the development of parthenogenetic porcine embryos.
本研究的主要目的是提高孤雌激活胚胎的发育率。在这项研究中，研究了三种抗氧化剂β-巯基乙醇（β-ME）、α-生育酚和细胞外超氧化物歧化酶（EC-SOD）对孤雌激活猪胚胎发育的抗氧化和抗凋亡作用。对照组孤雌激活猪胚胎发育到囊胚阶段的发育率为 8.1%；1μMβ-ME 组分别为 19.1%、14.6%和 5.0%；50μMα-生育酚组和 100μMα-生育酚组分别为 17.2%和 17.5%；EC-SOD 转基因小鼠胚胎成纤维细胞（Tg-MEF）和 EC-SOD 非转基因小鼠胚胎成纤维细胞（NTg-MEF）条件培养基组分别为 12.0%和 4.0%。β-ME、α-生育酚和 EC-SOD Tg-MEF 条件培养基均能提高孤雌激活猪胚胎的囊胚发育率（P<0.05）。在三种抗氧化剂的最佳条件下，分析了囊胚期总细胞和凋亡细胞的平均数量。与未添加抗氧化剂的对照组相比，三种抗氧化剂均增加了囊胚期的总细胞数量，降低了囊胚期的凋亡细胞数量（P<0.05）。当在 1μMβ-ME 和 100μMα-生育酚处理后的二细胞胚胎中检测到活性氧水平时，其水平低于对照组（P<0.05）。总之，发现三种抗氧化剂β-巯基乙醇、α-生育酚和 EC-SOD Tg-MEF 条件培养基在孤雌激活猪胚胎的发育中可以发挥强有力的刺激作用。
