Akademik Veri Yönetim Sistemi
Revue de Médecine Vétérinaire, cilt.170, sa.4/6, ss.87-94, 2019 (Scopus)
Percoll gradient and swim-up are the most common methods used to
obtain high quality intact sperm to be used for the in vitro fertilization
procedures. In this study, we evaluated the effects of swim-up and percoll
gradient methods on DNA integrity and other spermatological parameters.
To determine DNA breakdown, toluidine blue staining was used. Motility,
and progressive motility were analyzed via computer assisted semen
analyzer (CASA) system and the others (morphology, viability, membrane
and acrosome integrity) were evaluated via phase contrast and florescent
microscopy. The membrane integrity was tested by hypo-osmotic swelling
test. Also, the separated sperms were tested for in vitro embryos production.
Our results showed no differences between the two methods of separation
compared to the fresh samples in regard of sperm motility (P>0.05),
however, in the separated sperms with both methods, progressive motility
was higher than the fresh sperm and the highest results were obtained in
Percoll method (P<0.05). In comparison to swim-up, Percoll gradient
method maintained the highest normal sperm morphology rate and the
lowest acrosome defects (P<0.05). In addition, the membrane integrity,
intact acrosome and intact chromosome rates were also higher in the sperm
samples separated by percoll gradient method (P<0.05). The blastocyst rate
produced with sperm separated by Percoll gradient method was significantly
higher than the swim-up group (P<0.001). Sperm samples obtained after
Percoll gradient centrifugation could improve motility, morphological
intactness, and DNA integrity. Accordingly, it was concluded that the Percoll
gradient separation method is more useful then the swim-up for in vitro
sheep embryo production. Also, Toluidine blue staining could be a suitable
technique for determination of DNA integrity