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Comparison of the Motility, Morphology, and DNA Integrity of Cryopreserved Human Spermatozoa from Processing Semen before and after Cryopreservation

Rawiwan Jeamanukoolkit MD*, Chatchai Treetampinich MD*, Matchuporn Sukprasert MD*, Sasivimol Rattanasiri PhD**, Wicharn Choktanasiri MD*, Chonthicha Satirapod MD*

Affiliation : * Department of Obstetrics and Gynecology, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand ** Research Center, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand

Background : Cryopreservation of spermatozoa is an extremely important process in the field of male infertility. High quality preserved spermatozoa has been reported to retain better motility and DNA quality.
Objective : We showed the effect of semen preparation on the quality of cryopreservation of human spermatozoa. Material and Method: We compared cryopreserved spermatozoa separated by Sil-Select density gradient centrifugation before or after cryopreservation in terms of spermatozoa motility, morphology, and DNA integrity. Spermatozoa’s motility was determined by CASA; the morphology was determined by the eosin-methylene blue staining; and the DNA integrity was determined by TUNEL assay.
Results : We found that semen preparation before or after cryopreservation produced spermatozoa with comparable motility and morphology. However, semen preparation before cryopreservation showed a significantly higher total motile spermatozoa count (8.48 [2.47 to 37.87] vs. 5.35 [0.29 to 17.30], p-value <0.05) and significantly better DNA integrity as indicated by less DNA fragmentation (21.34±3.01 vs. 24.29±3.01, p<0.001).
Conclusion : We concluded that spermatozoa prepared by Sil-Select density gradient centrifugation before cryopreservation can improve quality of the preserved spermatozoa in terms of the total motile spermatozoa count and DNA integrity.

Keywords : Cryopreserved human spermatozoa, Sil-Select density gradient centrifugation, DNA integrity


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