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Two techniques for the preparation of boar spermatozoa for in vitro fertilization were studied: a simple washing procedure and centrifugation on a discontinuous Percoll gradient. Their respective effects on motility of spermatozoa were analysed by computer-assisted sperm analysis. The Percoll density gradient technique selected spermatozoa with significantly (P < 0.0001) enhanced motility and movement characteristics. In vitro matured oocytes inseminated with spermatozoa prepared by Percoll gradient centrifugation had significantly (P < 0.0001) greater cleavage rates than did oocytes inseminated with washed spermatozoa. This increased penetration ability was not due to an increased proportion of acrosome-reacted spermatozoa. Transmission electron microscopy revealed no unique ultrastructural differences between the spermatozoa from either preparation. Spermatozoa prepared by Percoll gradient centrifugation are recommended for insemination and studies of porcine in vitro fertilization.
