Advances in Urology / 2012 / Article / Tab 2

Review Article

Human Sperm Cryopreservation: Update on Techniques, Effect on DNA Integrity, and Implications for ART

Table 2

(a)–(c) Evaluation of DNA integrity after cryopreservation: description of the experimental design and conclusions.
(a)

AuthorsTest to evaluate DNA integrity Number of samplesCryopreservation method“Does the freezing-thawing procedure induce sperm DNA damage?”

Hamamah et al. [94]Acridine orange staining and Feulgen-DNA quantitative microspectrophotometry10UnspecifiedYes

Spanò et al. [44]SCSA + Acridine orange staining19Equilibration at 37°C, freezing in liquid nitrogen vapour at −80°C and then storage in liquid nitrogen at –196°CYes

Hammadeh et al. [95]Acridine orange staining59Computerized slow-stage freezer + static liquid nitrogen vapourYes

Donnelly et al. [6]COMET assay 40Equilibration at 37°C, freezing in liquid nitrogen vapour at −80°C and then storage in liquid nitrogen at –196°CYes

Gandini et al. [96]Acridine orange staining19Equilibration at 37°C, freezing in liquid nitrogen vapour at −80°C and then storage in liquid nitrogen at –196°CYes

de Paula et al. [40]TUNEL assay77: (i) 30 normozoospermic (ii) 47 oligozoospermic Use of freezer at –20°C, freezing in liquid nitrogen vapour, then storage in liquid nitrogen –196°CYes

Petyim and Choavaratana [43]Acridine orange staining 50Freezing with liquid nitrogen vapour + computerized program freezerYes

Nagamwuttiwong and Kunathikom [97]Acridine orange staining 20Freezing with liquid nitrogen vapourYes

Dejarkom and Kunathikom [98]Acridine orange staining 20Computerized controlled rate freezingYes

Thomson et al. [46]TUNEL assay60Use of programmable freezerYes

Thomson et al. [46]TUNEL assay 320Sample frozen with and without cryoprotectant by slow-controlled-rate method using a programmable freezerYes

Zribi et al. [45]TUNEL assay 15Equilibration at 37°C, freezing in liquid nitrogen vapour at −80°C, then storage in liquid nitrogen at –196°CYes

(b)

AuthorsTest to evaluate DNA integrityNumber of samplesCryopreservation technique“Does the freezing-thawing procedure induce sperm DNA damage?”

Donnelly et al. [6]COMET assay57: (i) 17 fertile (ii) 40 infertileEquilibration at 37°C, freezing in liquid nitrogen vapour at −80°C, then storage in liquid nitrogen at –196°CYes, but semen from fertile men appears to be more resistant to freezing damage

Kalthur et al. [49]COMET assay + Acridine orange staining44Equilibration at 37°C, static cooling at 4°C, cooling vapour phase, then storage in liquid nitrogen at –196°CYes, but morphologically abnormal sperms seems to be less resistant to freezing damage

Ahmad et al. [99]COMET assay196: (i) 30 normospermic (ii) 166 infertileFreezing with static-phase vapour cooling procedureYes, but the sperm DNA integrity of frozen samples of fertile men is higher

(c)

AuthorsTest to evaluate the DNA integrityNumber of samplesCryopreservation technique“Does the freezing-thawing procedure induce sperm DNA damage?”

Høst et al. [100]Immunoperoxidase detection of digoxigenin-labelled genomic DNA53: (i) 20 fertile (ii) 33 infertileConventional cryopreservationNo

Steele et al. [101]COMET assay21: (i) 9 control (ii) 12 with obstructive azoospermiaFreezing in liquid nitrogen vapourNo

Duru et al. [41]TUNEL assay + annexin V21Equilibration at 37°C, freezing in liquid nitrogen vapour at −80°C, then storage in liquid nitrogen at –196°CNo

Isachenko et al. [42]COMET assay 18Programmable slow freezing + vitrificationNo

Paasch et al. [50]TUNEL assay + flow cytometric kit for apoptosis84 Freezing at –20°C, freezing in liquid nitrogen vapor at –100°C, then storage in liquid nitrogen at –196°CNo