Author(s) (year) | Samples recruited | Conventional freezing procedure | Vitrification procedure | Comparison results |
---|---|---|---|---|
Saritha and Bongso [21] | 57 human semen samplesa | Glycerol used. RT 10 min, cryotube 0.85 mL, LN2 vapor, 15 + 15 min. Thawed RT 30–45 min. | Glycerol used. RT 10 min. Cryotube 0.85 mL. Plunged into LN2. Warmed RT 30–45 min. | No motility difference was found between two methods. |
Nawroth et al. [22] | 30 human semen samples. Native or swim-up. | Glycerol used. 0.25 mL straw, RT 10 min. 22 °C to 4 °C by − 5 °C/min; 4 °C to − 30 °C by − 10 °C/min; − 30 °C to − 140 °C by − 20 °C/min. Thawed in 37 °C water bath 50 s. | With or without permeable cryoprotectant. Copper loop, 20 μl or 0.25 mL straw. Plunged into LN2. Warmed in 37 °C medium 5–10 min. | Permeable cryoprotectant-free vitrification using copper loop resulted in higher motility with swim-up samples than conventional freezing. No difference in morphology. |
Chang et al. [23] | 30 healthy human semen samples | Freezing medium used. Biological freezer used. Thawing unspecified. | Freezing medium used. Plunged into LN2. Warming unspecified. | No difference in motility or DNA fragmentation was found between two methods. |
Vutyavanich et al. [24] | 30 normospermic human semen samples | Freezing medium used. 0.25 mL straw, RT 10 min. 20 °C to 5 °C by − 1 °C/min; − 5 °C to − 85 °C by − 10 °C/min. Thawed in 25–28 °C tap water. | Glycerol used. 0.25 mL straw, 4 °C 10 min. Plunged into LN2. Warmed in 25–28 °C tap water. | Vitrification gave superior motility and cryosurvival than conventional freezing. No difference in morphology or DNA integrity. |
Moskovtsev et al. [25] | 11 human semen samples. Washed. | Freezing medium used. LN2 vapor. Thawing unspecified. | Permeable cryoprotectant free. Warming unspecified. | Vitrification resulted in higher motility and progressive motility. |
Agha-Rahimi et al. [26] | 30 normozoospermic samples. Washed. | Glycerol used. Cryotube, LN2 vapor 30 min. Thawed in 37 °C water bath 10 min. | With or without glycerol. 30 μl drop, Plunged into LN2. Warmed at 37 °C medium 5–10 s. | Both methods resulted in similar motility, viability, recovery rate and DNA fragmentatioin. No permeable cryoprotectant was required for vitrification. |
Zhu et al. [27] | 58 human semen samples. Washed. | Glycerol used. RT 5 min. LN2 vapor 30 min. Cryogenic vial, 0.5 mL, LN2 vapor 30 min. Thawed in 37 °C water bath till melted. | Permeable cryoprotectant free. Cryogenic vial, 0.25 mL, RT 1 min. Plunged into LN2. Warmed in 42 °C water bath 1 min, 37 °C water bath till melted. | Vitrification with optimal sucrose concentration resulted in higher progressive motility, plasma membrane and acrosome integrity than conventional freezing. No differences in motility or DNA stability. |
Ali Mohamed [28] | 33human semen samples | Freezing medium used. 0.25 mL straw, RT 10 min, LN2 vapor 30 min. Thawed 37 °C water bath till melting. | permeable cryoprotectant free, 37 °C 5 min, 100 μl straw-in-straw, Plunged into LN2. Warmed in 42 °C medium. | Both methods had similar motility, viability and mitochondrial membrane potential. |
Slabbert et al. [20] | 35 human semen samples. Washed. | Freezing medium used. RT 10 min. 0.5 mL straw, LN2 vapor 15 min. Thawed in 23 °C 5 min. | permeable cryoprotectant free. 300 μl sample in 1.5 mL straw, RT 10 min. Plunged into LN2. Warmed in 42 °C medium 20 s. | Vitrification had higher mitochondrial membrane potential and lower percentage of DNA fragmentation than conventional freezing. No differences in motility. |
Tongdee et al. [29] | 37 normal human semen samples. Washed. | Freezing medium used. RT 10 min, Cryovial, 0.5 mL, 25 °C to 5 °C by − 1 °C/min; 5 °C to - 85 °C by − 10 °C/min. Thawed RT 15–20 min. | Freezing medium used. RT 10 min, Cryovial, 0.25 mL, plunged into LN2. Warmed RT 15–20 min. | Motility decreased more by vitrification. No difference in morphology or DNA intergrity between two methods. |
Aizpurua et al. [30] | 18 normozoospermic human semen samples. | Glycerol used, 1.8 mL tube, 4 °C 30 min, LN2 vapor 30 min. Thawed RT 30 min. | permeable cryoprotectant free, 37 °C 5 min, 20 μl drop, Plunged into LN2. Warmed at 37 °C 5 min. | vitrification had higher motility and normal morphology, and lower DNA fragmentation than conventional freezing. |
Karthikeyan et al. [31] | 20 severe oligoasthenozoospermia (SOA) and very SOA sampleb | Freezing medium used. 0.5 mL straw, − 85 °C in a deep freezer 1 h. Thawed at RT 10 min. | permeable cryoprotectant free. Cryologic with stripper. Warmed in 37 °C medium | Vitrification revealed higher motility vitality with very SOA samples than conventional freezeing. |
Le et al. [32] | 105 human semen samples. Washed and unwashed. | Glycerol used. Cryotube RT 10 min. LN2 vapor 15 min. Thawed at 37 °C water bath 5 min. | Glycerol used. RT 10 min. 30 μl drop, plunged into LN2. Warmed at 37 °C water bath 5 min. | Conventional freezing method resulted in higher motility, viability while vitrification resulted in higher normal morphology. |
Pabon et al. [33] | 47 human semen samples, Swim-up | Glycerol used. RT 10 min. 50 μl drop on dry ice, cryotube. Thawed RT 10 min, then 37 °C 10 min. | permeable cryoprotectant free. RT 3 min. Collector-grid, 5–10 μl drop, plunged to LN2. Warmed in 44 °C medium 3 min. | Vitrification presented higher motility, viability and mitochondrial activity than conventional freezing. |
Spis et al. [34] | 1 epididymal and 1 testicular sperm samples | Glycerol used. 20 + 8 capillaries. LN2 vapor 30 min. Thawed in 37 °C water bath 50 s. | permeable cryoprotectant free. 20 + 8 capillaries. Plunged into LN2. Warmed in 37 °C medium 20 s | Vitrification had higher mitochondrial membrane potential and motility in both epididymal and testicular capillaries than conventional freezingc. |