EFFECT OF OVIDUCTAL EPITHELIAL-CELLS ON FERTILIZATION OF PIG OOCYTES IN-VITRO
EFFECT OF OVIDUCTAL EPITHELIAL-CELLS ON FERTILIZATION OF PIG OOCYTES IN-VITRO
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DOI：
10.1016/0093-691x(94)90128-6
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发表时间：
1994-11-01
期刊：
影响因子：
2.8
通讯作者：
KATO, S
中科院分区：
文献类型：
--
作者：
KANO, K;MIYANO, T;KATO, S
猪卵母细胞与输卵管细胞共培养可降低多精受精的发生率。尚不清楚这种效应是否是由于分泌到培养基中的可溶性因子所致。本文研究了猪输卵管上皮细胞单层和细胞条件培养液对猪卵母细胞受精的影响。将体外成熟的猪卵母细胞与射出的公猪精子以1 × 10（5）或1 × 10（6）细胞/ml的浓度进行授精，并在5种培养系统之一中共培养：输卵管上皮细胞单层、成纤维细胞单层、输卵管上皮细胞条件培养基或成纤维细胞条件培养基，以及单独培养基（改良的TCM 199）。在所有5个系统中，大多数卵母细胞（范围85%至100%）被精子穿透。当卵母细胞与1 × 10（5）个细胞/ml的精子授精时，与输卵管上皮细胞和成纤维细胞共培养的卵母细胞中的单精子卵母细胞的百分比显著高于不含这些细胞的卵母细胞。相反，当卵母细胞与1 × 10（6）细胞/ml的精子授精时，与上皮细胞共培养的卵母细胞中的单精子卵母细胞的百分比显著高于与成纤维细胞和对照培养基中培养的卵母细胞。在输卵管上皮细胞条件培养基中，当卵母细胞与1 × 10（5）和1 × 10（6）个细胞/ml的精子授精时，观察到对多精受精的抑制作用。在成纤维细胞条件培养基中不存在这种效应。此外，上皮细胞的作用在培养期间保持不变，而与成纤维细胞共培养的单精子卵母细胞的比例逐渐下降，16 h后为0%。这些结果表明，来自输卵管上皮细胞的可溶性因子减少了穿透卵母细胞的精子数量，但不抑制高受精率。
The incidence of polyspermy is reduced by co-culture of pig oocytes with oviductal cells. It is not known whether the effect is due to soluble factors secreted into the medium. Oviductal epithelial cell monolayers and cell-conditioned media were prepared and their effects on fertilization of pig oocytes were examined. In vitro matured pig oocytes were inseminated with ejaculated boar spermatozoa at a concentration of 1 x 10(5) or 1 x 10(6) cells/ml and co-cultured in one of 5 culture systems: an oviductal epithelial cell monolayer, a fibroblast monolayer, an oviductal epithelial cell-conditioned medium, .or a fibroblast-conditioned medium, and medium alone(modified-TCM199). In all 5 systems, the majority (range 85 to 100%) of the oocytes were penetrated by sperm, When oocytes were inseminated with spermatozoa at a concentration of 1 x 10(5) cells/ml, the percentages of monospermic oocytes were significantly higher in the oocytes co-cultured with oviductal epithelial cells and fibroblasts than that of the oocytes cultured without these cells. In contrast, when oocytes were inseminated with spermatozoa at a concentration of 1 x 10(6) cells/ml, the percentages of monospermic oocytes were significantly higher in the oocytes co-cultured with epithelial cells than those cultured with the fibroblasts and in the control medium. The suppressive effect on polyspermy was observed in the oviductal epithelial cells-conditioned medium when oocytes were inseminated with spermatozoa at both concentrations of 1 x 10(5) and 1 x 10(6) cells/ml. The effect was absent in the fibroblasts-conditioned medium. Moreover, the effect of the epithelial cells was maintained during the culture period, whereas the proportion of monospermic oocytes cocultured with fibroblasts showed a gradual decrease, reaching 0% after 16 h. These results suggest that a soluble factor(s) derived from the oviductal epithelial cells decreased the number of spermatozoa penetrating the oocytes without suppressing the high rate of fertilization.