Vitrification and conventional freezing methods in sperm cryopreservation: A systematic review and meta-analysis

The objective is to systematically review and synthesize the literature on the efficacy with two different cryopreservation methods used for human spermatozoa and evaluate whether vitrification protocol and quality of sperm influence effect estimates.

The following electronic databases were searched up to September 2017: Pubmed, Embase and Web of Science. The search strategy used the following the relevant medical subject heading (MeSH) terms, keywords, and word variants for: sperm parameters, conventional freezing, and vitrification. Queries were limited to those involving humans. Randomized controlled trials (RCTs) that published in English languages were considered eligible. Studies and references were included if they reported total motility, progressive motility, morphology, or DNA fragmentation index (DFI) for vitrified or conventional cryopreserved human spermatozoa. Patients recruited in RCTs considering sperm vitrification as one of the experimental arms and conventional freezing (including slow freezing or vapor fast freezing) sperm control as the other. Studies that had high risks of allocation concealment were excluded when performing sensitivity analysis. We specified 2 subgroup variables, including vitrification protocol and quality of spermatozoa cryopreserved, to investigate sources of heterogeneity. A meta-analysis was performed using a random effects (I² > 50%) or fixed effects (I² < 50%) model to calculate weighted mean differences (MD) and 95% CI.

The search yielded a total of 2428 articles and 13 RCTs were included for analysis. They involved 486 vitrified and 486 conventional cryopreserved sperm samples. Four sperm parameters were reported as mean differences and based on adjusted estimates in all included studies. Meta-analysis of these studies showed significantly higher total motility [weighted mean differences (WMD) 6.98; 95% confidence interval (CI) 2.94; 11.02; P < 0.0001] and progressive motility [WMD 4.59; 95% CI 0.78; 8.39; P = 0.02] of past-thawed sperm following vitrification compared with conventional freezing methods. However, DNA fragmentation index (DFI) [WMD -1.18; 95% CI -2.81; 0.45; P = 0.16] and morphology [WMD 0.11; 95% CI -0.42; 0.63; P = 0.69] of past-thawed sperm are similar between two freezing groups. Subgroup analysis shown that the vitrification protocol and quality of spermatozoa are potential risk factors for the efficacy of vitrification. Higher past-thawed sperm parameters following the cryoprotectants-free (CPAs-free) vitrification were observed, as well as a lower past-thawed sperm parameters with the cryoprotectants-presence (CPAs-presence) vitrification, which could reflect the CPAs related cytotoxicity. Meanwhile, vitrification had higher ability in preservation of high quality of spermatozoa compared with vitrification of low quality spermatozoa.

According to the results of present meta-analysis, vitrification is superior to conventional freezing methods in preservation of spermatozoa, regarding total and progressive motility. However, the efficacy of vitrification is influence by using different vitrification protocol and cryopreservation of different quality spermatozoa. It is must emphasized that the results of present meta-analysis is limited by the small number of studies of variable vitrification protocol. Further well conducted studies are required to confirm the efficacy of vitrification in cryopreservation of spermatozoa, in addition, allow the examination of the two cryopreservation methods in terms of pregnancy achievement and determination of the role of clinical variable on efficacy of vitrification.