Wongsrikeao P, Kaneshige Y, Ooki R, Taniguchi M, Agung B, Nii M, Otoi T
Laboratory of Animal Reproduction, Department of Veterinary Science, Yamaguchi University, Yoshida, Yamaguchi, Japan.
Reprod Domest Anim. 2005 Apr;40(2):166-70. doi: 10.1111/j.1439-0531.2005.00576.x.
The present study was conducted to investigate the effects of attachment of cumulus cells to porcine oocytes during the process of maturation and fertilization on the nuclear maturation, fertilization and subsequent development after in vitro fertilization (IVF). In the first experiment, the cumulus cells were removed from cumulus-oocyte complexes (COCs) at 0, 24 and 42 h after the onset of maturation culture and were then cultured until reaching 42 h of cultivation. In the second experiment, COCs were denuded as described in the first experiment, then fertilized and cultured for 7 days. As a control, cumulus cells were allowed to maintain attachment to the oocytes until the end of IVF. The proportion of oocytes reaching metaphase II significantly increased with the delay in the removal treatment of cumulus cells. The proportion of normal fertilization gradually increased with delay in the removal treatment of cumulus cells from COCs until the end of IVF. However, no significant difference in the proportion of normal fertilization was found between the 42-h and control groups. The removal treatment of cumulus cells in the 0- and 24-h group significantly (p < 0.05) decreased the proportion of cleaved embryos when compared with the control, and none of them developed to the blastocyst stage. The proportion of development to the blastocyst stage was significantly higher (p < 0.05) in the control group than in the 42-h group (18.1% vs 12.4%; p < 0.05). The present study indicates that the attachment of cumulus cells to the oocyte during maturation and fertilization is important to support oocyte nuclear maturation, fertilization and subsequent embryo development. Particularly, the attachment of cumulus cells to the oocyte during IVF promotes embryonic development.
本研究旨在探讨卵丘细胞在成熟和受精过程中与猪卵母细胞的附着对体外受精(IVF)后核成熟、受精及后续发育的影响。在第一个实验中,在成熟培养开始后的0、24和42小时从卵丘 - 卵母细胞复合体(COCs)中去除卵丘细胞,然后培养至42小时。在第二个实验中,COCs按第一个实验所述去除卵丘细胞,然后受精并培养7天。作为对照,卵丘细胞在IVF结束前一直保持与卵母细胞的附着。随着卵丘细胞去除处理的延迟,达到减数分裂中期II的卵母细胞比例显著增加。从COCs中去除卵丘细胞的处理延迟至IVF结束时,正常受精比例逐渐增加。然而,42小时组与对照组之间在正常受精比例上未发现显著差异。与对照组相比,0小时和24小时组去除卵丘细胞的处理显著(p < 0.05)降低了卵裂胚胎的比例,且它们均未发育至囊胚阶段。对照组发育至囊胚阶段的比例显著高于42小时组(18.1%对12.4%;p < 0.05)。本研究表明,卵丘细胞在成熟和受精过程中与卵母细胞的附着对于支持卵母细胞核成熟、受精及后续胚胎发育至关重要。特别是,IVF期间卵丘细胞与卵母细胞的附着促进胚胎发育。