Bijttebier J, Van Soom A, Meyer E, Mateusen B, Maes D
Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.
Theriogenology. 2008 Sep 1;70(4):715-24. doi: 10.1016/j.theriogenology.2008.04.046. Epub 2008 Jun 24.
Porcine follicular fluid (pFF), as a supplement of maturation media, has been shown several times to improve the in vitro production (IVP) of porcine embryos. As a transudate of serum, pFF contains locally produced factors in addition to the ones derived from serum. The objective of this study was to determine the additional positive effects of these pFF specific factors on the nuclear and cytoplasmic maturation of porcine oocytes. Follicular fluid and autologous serum were collected from sows in the preovulatory phase of the estrous cycle. Subsequently, oocytes from prepubertal gilts were matured in NCSU23 supplemented with either 10% pFF or 10% autologous serum derived from the same sow. Oocytes were then fertilized and the putative zygotes were cultured for 7 days. Nuclear maturation and cumulus expansion were assessed after the maturation culture. For evaluation of cytoplasmic maturation, oocyte glutathione (GSH) content, fertilization parameters and embryonic development were evaluated. After in vitro maturation (IVM) of the oocytes, both cumulus expansion rate and oocyte GSH content were increased for oocytes matured in pFF (P<0.05). More monospermic penetration was found when cumulus-intact oocytes had been matured in 10% pFF but this effect was lost after fertilization of cumulus denuded oocytes indicating that the pFF was acting through the cumulus. We speculate that the increased cumulus expansion and increased glutathione content, which were prevalent after IVM in pFF, are responsible for the positive effects on fertilization and the pre-implantation development of the embryos.
猪卵泡液(pFF)作为成熟培养基的补充物,已多次被证明可提高猪胚胎的体外生产(IVP)。作为血清的渗出液,pFF除了含有源自血清的因子外,还含有局部产生的因子。本研究的目的是确定这些pFF特异性因子对猪卵母细胞核成熟和细胞质成熟的额外积极作用。在发情周期的排卵前期从母猪收集卵泡液和自体血清。随后,将青春期前后备母猪的卵母细胞在补充有10% pFF或来自同一头母猪的10%自体血清的NCSU23中成熟。然后使卵母细胞受精,并将假定的合子培养7天。成熟培养后评估核成熟和卵丘扩展。为了评估细胞质成熟,评估了卵母细胞谷胱甘肽(GSH)含量、受精参数和胚胎发育。卵母细胞体外成熟(IVM)后,在pFF中成熟的卵母细胞的卵丘扩展率和卵母细胞GSH含量均增加(P<0.05)。当完整卵丘卵母细胞在10% pFF中成熟时,发现更多的单精子穿透,但在去除卵丘的卵母细胞受精后这种效果消失,这表明pFF是通过卵丘起作用的。我们推测,pFF中IVM后普遍存在的卵丘扩展增加和谷胱甘肽含量增加,是对受精和胚胎植入前发育产生积极影响的原因。