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

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.

Thinning and drilling laser-assisted hatching in thawed embryo transfer: A randomized controlled trial.

OBJECTIVE: In frozen and thawed embryos, the zona pellucida (ZP) can be damaged due to hardening. Laser-assisted hatching (LAH) of embryos can increase the pregnancy rate. This study compared thinning and drilling of the ZP before frozen embryo transfer (FET). METHODS: Patients were randomly allocated into two groups for LAH using thinning or drilling on day 2 after thawing. Twenty-five percent of the ZP circumference and 50% of the ZP thickness was removed in the thinning group, and a hole 40 µm in diameter was made in the drilling group. RESULTS: A total of 171 in vitro fertilization/intracytoplasmic sperm injection FET cycles, including 85 cycles with drilling LAH and 86 cycles with thinning LAH, were carried out. The thinning group had a similar ß-human chorionic gonadotropin-positive rate (38.4% vs. 29.4%), implantation rate (16.5% vs. 14.4%), clinical pregnancy rate (36.0% vs. 25.9%), miscarriage rate (5.8% vs. 2.4%), ongoing pregnancy rate (30.2% vs. 23.5%), and multiple pregnancy rate (7.0% vs. 10.6%) to the drilling LAH group. There were no significant differences in pregnancy outcomes between subgroups defined based on age (older or younger than 35 years) or ZP thickness (greater or less than 17 µm) according to the LAH method. CONCLUSION: The present study demonstrated that partial ZP thinning or drilling resulted in similar outcomes in implantation and pregnancy rates using thawed embryos, irrespective of women's age or ZP thickness.