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Volume 7, Issue 1 (January 2005) 7, 86–92; 10.1111/j.1745-7262.2005.00008.x

Comparative study on density gradients and swim-up preparation techniques utilizing neat and cryopreserved spermatozoa

Shyam S R Allamaneni, Ashok Agarwal, Sreedhar Rama, Pavithra Ranganathan and Rakesh K Sharma

Shyam S R Allamaneni, Ashok Agarwal, Sreedhar Rama, Pavithra Ranganathan and Rakesh K Sharma

Advance online publication 1 January 2005


Aim: To 1) compare post-wash and post-thaw parameters of sperm processed with PureSperm density gradient technique and swim-up method; and 2) test the efficacy of two commonly available density gradient media PureSperm and ISolate.

Methods: This prospective study used semen specimens from 22 patients. Specimens from nine patients were processed by both PureSperm density gradient and swim-up method. These specimens were then cryopreserved. Thirteen specimens were processed by both PureSperm (40 % and 80 %) and Isolate (50 % and 90 %) double density gradient techniques. The two fractions processed by both PureSperm and swim-up were analyzed for post-wash sperm characteristics. Post-thaw analysis was done after 24 hours. Sperm fractions obtained after processing with PureSperm and ISolate were compared for post-wash sperm characteristics and ROS levels.

Results: Specimens prepared with PureSperm had significantly higher median total motile sperm counts (TMSC) (32.2×106 vs. 17.6×106), recovery rates (69.2 %vs. 50.0 %), and longevity at 4 hours (83.0 %vs. 55.0 %) compared to specimen prepared by swim-up. Post-thaw specimens also had a higher recovery and longevity at 4 hours with PureSperm as compared to the swim-up. Semen specimens processed by PureSperm had significantly higher total sperm count, TMSC, and percentage recovery rates (30.0 %vs. 19.7 %) than ISolate.

Conclusion: Semen quality is better preserved in fresh and cryopreserved semen prepared with PureSperm density gradient compared to swim-up. A significant enrichment of sperm is observed with PureSperm compared to ISolate. Higher recovery rates of mature motile sperm obtained after PureSperm sperm preparation may be beneficial for successful ART.

Keywords: spermatozoa, cryopreservation, density gradient centrifugation, sperm motility

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