In-vitro penetration of pig oocytes matured in culture by frozen-thawed ejaculated spermatozoa

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Abstract
Summary. 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 × 106 spermatozoa/ml. Wide variation in penetration rates (16–89%) was observed in oocytes inseminated in medium containing 5mm caffeine and at 25–50 × 106 spermatozoa/ml obtained from 6 boars, regardless of sperm motility. At 25–50 × 106 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 μg/ml. When heparin was included in the medium with 5mm caffeine, it inhibited the efficacy of caffeine to promote sperm penetration of oocytes. Keywords: in-vitro fertilization; oocyte; pig; frozen ejaculated sperm

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