Wang W H, Niwa K, Okuda K
Division of Animal Science, Faculty of Agriculture, Okayama University, Japan.
J Reprod Fertil. 1991 Nov;93(2):491-6. doi: 10.1530/jrf.0.0930491.
Pig oocytes matured in culture were inseminated with frozen-thawed ejaculated spermatozoa without preincubation in modified tissue culture medium (TCM) 199. High penetration rates (85-89%) and increased incidence of polyspermy were obtained at 25-100 x 10(6) spermatozoa/ml. Wide variation in penetration rates (16-89%) was observed in oocytes inseminated in medium containing 5mM caffeine and at 25-50 x 10(6) spermatozoa/ml obtained from 6 boars, regardless of sperm motility. At 25-50 x 10(6) spermatozoa/ml, penetration rates of oocytes were dependent upon the concentration of caffeine in the medium: there was no penetration without caffeine, but penetration was highest (89%) with 5mM caffeine. None of the oocytes was penetrated in the medium supplemented with heparin at 5-40 micrograms/ml. When heparin was included in the medium with 5mM caffeine, it inhibited the efficacy of caffeine to promote sperm penetration of oocytes.
将在体外成熟的猪卵母细胞与经冷冻解冻的射出精子进行受精，精子无需在改良的组织培养液（TCM）199中预孵育。当精子浓度为25 - 100×10⁶/ml时，可获得较高的穿透率（85 - 89%），但多精入卵的发生率也有所增加。在含有5mM咖啡因的培养液中，当精子浓度为25 - 50×10⁶/ml且取自6头公猪时，无论精子活力如何，卵母细胞的穿透率都有很大差异（16 - 89%）。当精子浓度为25 - 50×10⁶/ml时，卵母细胞的穿透率取决于培养液中咖啡因的浓度：无咖啡因时无穿透发生，但在含5mM咖啡因时穿透率最高（89%）。在添加了5 - 40μg/ml肝素的培养液中，没有卵母细胞被穿透。当在含5mM咖啡因的培养液中加入肝素时，它会抑制咖啡因促进精子穿透卵母细胞的效果。
