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Centrifugation on Percoll density gradient enhances motility, membrane integrity and in vitro fertilizing ability of frozen–thawed boar sperm

  • Michiko Noguchi (a1), Koji Yoshioka (a2), Hirokazu Hikono (a1), Gentaro Iwagami (a3), Chie Suzuki (a1) and Kazuhiro Kikuchi (a4)...


The effects of Percoll density gradient centrifugation on sperm quality, in vitro fertilizability and developmental capacity of frozen–thawed boar sperm were evaluated. Two-step density gradient centrifugation by Percoll enhanced significantly the motility parameters of sperm compared with a simple centrifugation procedure. Percentages of motile sperm and sperm with intact plasma and acrosome membranes after Percoll separation were significantly greater than those after simple centrifugation. The rates of penetration, cleavage and blastocyst formation after in vitro fertilization were significantly improved by Percoll separation compared with simple centrifugation and were influenced positively by the intactness of sperm head membranes, but not any sperm motility parameters. However, insemination with increased concentrations of sperm prepared by Percoll gradient centrifugation did not improve the success of fertilization and embryo development in vitro. Our results indicate that the integrity of sperm head membranes after Percoll separation is important for successful embryo development in vitro, more so than sperm motility.


Corresponding author

All correspondence to: Koji Yoshioka. National Institute of Animal Health, Kannondai 3–1-5, Tsukuba, Ibaraki 305–0856, Japan. Tel: +81 29 838 7784. Fax: +81 29 838 7880. e-mail:


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