Cryopreservation of human spermatozoa by vitrification versus conventional rapid freezing: Effects on motility, viability, morphology and cellular defects

Abstract

Objectives: Sperm cryopreservation has great potential for male infertility treatment as used in assisted reproduction technology (ART). There are a variety of cryopreservation methods in order to preserve sperm in a long term. Although conventional freezing and vitrification now are used widely, they have damage on sperm parameters as well as sperm DNA integrity. It is necessary to answer which method is better and appropriate for sperm cryopreservation. The aim of this study was to compare the effect of conventional freezing and vitrification regarding to motility, vitality and morphology of sperm found in washed and unwashed samples.

Study design: One hundred and five human fresh semen samples were divided into washed and unwashed halves using density-gradient centrifugation. Each group then was split into two aliquots: one cryopreserved by conventional freezing and the other by vitrification, using SpermFreeze Solution™ (Vitrolife, Västra Frölunda, Sweden) containing glycerol as a cryoprotectant. The sperm parameters were analyzed and compared between six groups: washed fresh (FW), unwashed fresh (FU), washed conventional freezing (CfW), unwashed conventional freezing (CfU), washed vitrification (VitW) and unwashed vitrification (VitU) samples.

Results: Sperm progressive motility, vitality and normal morphology significantly decreased, together with an appreciable increase in sperm head, midpiece and tail defects when comparing to the fresh sperm parameters after thawing in all groups. In conventional freezing method groups, progressive motility and vitality were substantially higher than that in vitrification method groups. However, vitrification gave better results in normal morphology rates. Additionally, sperm head, midpiece and tail defects were significant lower in two vitrification groups in comparison with conventional freezing groups. Interestingly, washed groups had better sperm parameters than unwashed groups so that washing process before frozen seemed to improve sperm parameters.

Conclusion: Conventional freezing method resulted in better motility, viability in both washed/unwashed groups. On the contrary, spermatozoa undergoing vitrification were healthier regarding morphology with less defects than conventional freezing. Sperm washing before frozen was a beneficial preparation to sperm cryopreservation.

Keywords: Conventional freezing; Cryopreservation; Human sperm; Vitrification.