Muzikova E, Clark D A
Department of Medicine, McMaster University, Hamilton, Ontario, Canada.
Hum Reprod. 1995 May;10(5):1172-7. doi: 10.1093/oxfordjournals.humrep.a136113.
This study was designed to investigate whether the polyamines spermine, spermidine and putrescine can prevent the arrest of development of in-vitro fertilized murine oocytes from CD1 strain mice. Development in M16 or Brinster's medium with or without polyamine supplementation was assessed and the ability of blastocysts to attach and grow out on laminin was compared with in-vivo-generated embryo blastocysts. Oocytes from old (20 weeks) females showed lower rates of fertilization and subsequent development in comparison with oocytes from young (6-8 weeks old) females, but in both cases < 5% of in-vitro fertilized oocytes achieved the blastocyst stage. In-vitro fertilized young oocytes showed enhanced development to the blastocyst stage with the addition of the polyamine putrescine, although lesser enhancement was seen with spermine and spermidine; this effect was only evident when the oocytes were grown in Brinster's medium, reflecting the need for a particular set of culture conditions for growth-enhancing agents to be effective. Blastocysts generated in putrescine-supplemented Brinster's medium were able to implant on laminin-coated surfaces in vitro and to respond to the cytokines colony-stimulating factor-1, epidermal growth factor and platelet-derived growth factor with increased trophoblast outgrowth to a greater degree than in-vivo-generated blastocysts. Murine in-vitro fertilized embryo development was not enhanced by co-culture with Vero cells. Putrescine was the most effective polyamine, enhancing the in-vitro fertilized oocyte development to blastocysts in a permissive environment.
本研究旨在调查多胺精胺、亚精胺和腐胺是否能防止来自CD1品系小鼠的体外受精鼠卵母细胞发育停滞。评估了在添加或不添加多胺的M16或布林斯特培养基中的发育情况，并将囊胚在层粘连蛋白上附着和生长的能力与体内产生的胚胎囊胚进行了比较。与年轻（6 - 8周龄）雌性小鼠的卵母细胞相比，老龄（20周）雌性小鼠的卵母细胞受精率和随后的发育率较低，但在这两种情况下，体外受精的卵母细胞中只有不到5%能发育到囊胚阶段。添加多胺腐胺后，体外受精的年轻卵母细胞发育到囊胚阶段的能力增强，不过精胺和亚精胺的增强作用较小；这种效应只有当卵母细胞在布林斯特培养基中培养时才明显，这反映了生长促进剂发挥作用需要特定的一组培养条件。在添加腐胺的布林斯特培养基中产生的囊胚能够在体外层粘连蛋白包被的表面着床，并对细胞因子集落刺激因子-1、表皮生长因子和血小板衍生生长因子作出反应，其滋养层生长的增加程度大于体内产生的囊胚。与Vero细胞共培养并未增强小鼠体外受精胚胎的发育。腐胺是最有效的多胺，在适宜的环境中可增强体外受精卵母细胞发育成囊胚的能力。
