de Matos D G, Furnus C C, Moses D F, Baldassarre H
Fundación Margarita Pérez Companc, Centro de Investigaciones Reproductivas Pérez Companc, Buenos Aires, Argentina.
Mol Reprod Dev. 1995 Dec;42(4):432-6. doi: 10.1002/mrd.1080420409.
The present study was carried out to evaluate if the addition of cysteamine to the culture medium during in vitro maturation of bovine oocytes increased the glutathione (GSH) levels in the mature oocytes, and if these changes may promote an improvement on in vitro development to the blastocyst stage. Follicular oocytes from slaughterhouse ovaries were matured in TCM 199 supplemented with 10% (v/v) fetal calf serum, hormones, and O (control), 25, 50, or 100 muM of cysteamine for 24 hr. After in vitro maturation the oocytes were fertilized and cultured for 8 days. The percentage of embryos that developed to the blastocyst stage was significantly higher (P < 0.01) for oocytes matured in medium containing 100 muM of cysteamine than for those matured in control medium. Moreover, the intracellular GSH levels were increased (P < 0.05) in oocytes matured with 100 muM of cysteamine with respect to control. No differences were observed in maturation and cleavage rates, and in the mean cell numbers per blastocyst among treatments (P > 0.05). These results indicate that the addition of thiol compounds such as cysteamine to maturation medium increases the efficiency of in vitro blastocyst production from immature bovine oocytes. The higher levels of GSH in oocytes matured in the presence of cysteamine suggest that the beneficial effects of cysteamine on in vitro maturation and subsequent development after in vitro fertilization are mediated by GSH.
本研究旨在评估在牛卵母细胞体外成熟培养过程中添加半胱胺是否会增加成熟卵母细胞中谷胱甘肽（GSH）的水平，以及这些变化是否会促进体外发育至囊胚阶段。从屠宰场卵巢采集的卵泡卵母细胞在添加10%（v/v）胎牛血清、激素以及0（对照）、25、50或100μM半胱胺的TCM 199中成熟培养24小时。体外成熟后，卵母细胞进行受精并培养8天。在含有100μM半胱胺的培养基中成熟的卵母细胞发育至囊胚阶段的胚胎百分比显著高于对照培养基中成熟的卵母细胞（P<0.01）。此外，与对照相比，用100μM半胱胺成熟的卵母细胞内GSH水平升高（P<0.05）。各处理间在成熟率、卵裂率以及每个囊胚的平均细胞数方面均未观察到差异（P>0.05）。这些结果表明，在成熟培养基中添加诸如半胱胺之类的硫醇化合物可提高未成熟牛卵母细胞体外产生囊胚的效率。在半胱胺存在下成熟的卵母细胞中较高的GSH水平表明，半胱胺对体外成熟及体外受精后后续发育的有益作用是由GSH介导的。
