Takahashi M, Nagai T, Hamano S, Kuwayama M, Okamura N, Okano A
Department of Animal Reproduction, National Institute of Animal Industry, Ibaraki, Japan.
Biol Reprod. 1993 Aug;49(2):228-32. doi: 10.1095/biolreprod49.2.228.
The purpose of this investigation was to determine the effect of beta-mercaptoethanol (beta-ME) and cysteamine, low-molecular-weight thiol compounds, on the development and intracellular glutathione content of bovine embryos obtained by in vitro fertilization of in vitro-matured oocytes. Embryos developed to the 6-8-cell stage after in vitro fertilization were cultured without feeder cells in TCM-199 containing 10% fetal calf serum with or without beta-ME or cysteamine. The percentage of embryos that developed to the blastocyst and hatched blastocyst stages were significantly higher in medium containing beta-ME or cysteamine. Also, total intracellular glutathione levels were higher for embryos cultured in the medium with beta-ME or cysteamine than for those cultured in medium without thiol compounds. Moreover, when buthionine sulfoximine, a specific inhibitor of glutathione synthesis, was added to medium containing thiol compounds, there was a reduction both in the development of embryos to the blastocyst stage and in intracellular glutathione content. These results indicate that the inclusion of low-molecular-weight thiol compounds aids the in vitro development of bovine embryos without feeder cells and that the effect of thiol compounds is mediated through the increase of intracellular glutathione levels.
本研究的目的是确定β-巯基乙醇（β-ME）和半胱胺（一种低分子量硫醇化合物）对通过体外成熟卵母细胞体外受精获得的牛胚胎发育及细胞内谷胱甘肽含量的影响。体外受精后发育至6-8细胞阶段的胚胎，在含有10%胎牛血清且添加或不添加β-ME或半胱胺的TCM-199中，无饲养层细胞培养。在含有β-ME或半胱胺的培养基中，发育至囊胚和孵化囊胚阶段的胚胎百分比显著更高。此外，在含有β-ME或半胱胺的培养基中培养的胚胎，其细胞内谷胱甘肽总水平高于在无硫醇化合物培养基中培养的胚胎。而且，当向含有硫醇化合物的培养基中添加谷胱甘肽合成的特异性抑制剂丁硫氨酸亚砜胺时，胚胎发育至囊胚阶段的比例以及细胞内谷胱甘肽含量均降低。这些结果表明，添加低分子量硫醇化合物有助于无饲养层细胞时牛胚胎的体外发育，且硫醇化合物的作用是通过增加细胞内谷胱甘肽水平介导的。
