Process and Pitfalls of Sperm Cryopreservation

Rozati, Hamoun; Handley, Thomas J.; Jayasena, Channa N.

doi:10.3390/jcm6090089

Cited by 41 publications

(22 citation statements)

References 108 publications

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“…These damages can reduce fertility of post-thawed spermatozoa [8]. However, the basic factor limiting the effectiveness of cryopreservation process is the initial quality of the obtained spermatozoa [9]. Samples containing large number of functionally and morphologically impaired and dead spermatozoa display a lower tolerance of the cryopreservation process than those characterized by good initial quality [10].…”

Section: Introductionmentioning

confidence: 99%

The influence of Percoll® density gradient centrifugation before cryopreservation on the quality of frozen wisent (Bison bonasus) epididymal spermatozoa

et al. 2022

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Background The wisent (Bison bonasus) is a species that has undergone a population bottleneck. Homozygosity is prevalent within the population and may have a negative impact on semen quality in wisent bulls. Semen samples containing a large amount of functionally and morphologically impaired or dead spermatozoa have lower tolerance for cryopreservation process. Such samples are prone to involve damage acrosomes, to produce and release reactive oxygen which negatively affects proper function of spermatozoas. It is a good practice to select intact and viable gametes before subjecting the sample to cryopreservation to improve the efficiency of this process. The aim of this study was to assess the ability of Percoll® density gradient centrifugation in order to improve the quality of wisent spermatozoa after cryopreservation. Spermatozoa samples were analysed with computer-assisted semen analysis system and flow cytometry. Results Percoll® density gradient centrifugation resulted in increased percentage of motile spermatozoa, higher proportion of spermatozoa with normal morphology and proper functionality but also in a significant reduction of the total number of gametes. Nevertheless, the concentration of frozen spermatoza was still sufficient for obtaining a few complete insemination doses suggested for cattle from each epididymis. Conclusions While creating a high-quality genetic reserve, for in vitro fertilisation purposes, eliminating detritus and improving the overall quality of samples is more important than total number of spermatozoa. For these reasons, the achievement of higher post thaw quality of spermatozoa justifies the purification of samples by centrifugation in a Percoll® density gradient prior to the cryopreservation process.

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Section: Introductionmentioning

confidence: 99%

The influence of Percoll® density gradient centrifugation before cryopreservation on the quality of frozen wisent (Bison bonasus) epididymal spermatozoa

et al. 2022

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“…Use of frozen-thawed (FT) spermatozoa is currently the best way to (i) overcome some infertility issues in men [1,2], (ii) accelerate genetic progress in livestock species [3], and (iii) contribute to the survival of endangered species [4]. Despite improvements made in cryopreservation procedures and its inherent value, FT-semen is yet not routinely used neither for most livestock species [5,6] nor humans [7,8]. The main drawback of FT is the lower fertility rates after artificial insemination (AI), compared to natural mating or AI with unfrozen semen [9,10].…”

Section: Introductionmentioning

confidence: 99%

Cryopreservation Differentially Alters the Proteome of Epididymal and Ejaculated Pig Spermatozoa

Pérez-Patiño

Barranco

et al. 2019

IJMS

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Cryopreservation induces differential remodeling of the proteome in mammalian spermatozoa. How these proteome changes relate to the loss of sperm function during cryopreservation remains unsolved. The present study aimed to clarify this issue evaluating differential changes in the proteome of fresh and frozen-thawed pig spermatozoa retrieved from the cauda epididymis and the ejaculate of the same boars, with clear differences in cryotolerance. Spermatozoa were collected from 10 healthy, sexually mature, and fertile boars, and cryopreserved using a standard 0.5 mL-straw protocol. Total and progressive motility, viability, and mitochondria membrane potential were higher and membrane fluidity and reactive oxygen species generation lower in frozen-thawed (FT) epididymal than ejaculated spermatozoa. Quantitative proteomics of fresh and FT spermatozoa were analyzed using a LC-ESI-MS/MS-based Sequential Window Acquisition of All Theoretical Spectra approach. Cryopreservation quantitatively altered more proteins in ejaculated than cauda epididymal spermatozoa. Differential protein–protein networks highlighted a set of proteins quantitatively altered in ejaculated spermatozoa, directly involved in mitochondrial functionality which would explain why ejaculated spermatozoa deteriorate during cryopreservation.

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“…Sperm cryopreservation is the only available method of preserving male reproductive potential, and has been used widely since the 1970s to treat couples with infertility [ 1 ]. Sperm banking has become an important part of any assisted reproductive technology (ART) program, and is essential for preserving fertility in male patients with cancer and some autoimmune diseases [ 2 ].…”

Section: Introductionmentioning

confidence: 99%

Increased cryo-survival rate in ejaculated human sperm from infertile men following pre-freeze in vitro myo-inositol supplementation

Saleh

Assaf

Maged

et al. 2018

Clin Exp Reprod Med

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ObjectiveTo investigate the effects of in vitro myo-inositol (Myo-Ins) supplementation of cryopreserved human semen on the cryo-survival rate (CSR).MethodsSemen samples were obtained from 41 infertile men. Following routine semen analysis, each sample was divided into two equal aliquots (0.5 mL each). One aliquot was treated with 1 mg of Myo-Ins dissolved in 10 µL of sperm preparation medium. The second aliquot was treated with 10 µL of the same medium (control). Both aliquots were incubated for 20 minutes prior to freezing to slow the freezing process. The frozen samples were examined for post-thaw percentages of total motility (TM), progressive motility (PM), and the CSR, defined as the percentage of post-thaw TM divided by the percentage of pre-freeze TM and multiplied in 100. The results were expressed as median and interquartile range (25th and 75th percentiles).ResultsThe pre-freeze TM (50% [30%–50%]) and PM (35% [20%–35%]) were significantly higher than the post-thaw TM and PM in the Myo-Ins group (15% [10%–35%] and 10% [5%–20%]; p<0.001 and p<0.001, respectively) and the control group (10% [6%–30%] and 5% [3%–15%]; p<0.001 and p<0.001, respectively). The CSR of the 41 semen aliquots supplemented with Myo-Ins (40% [25%–70%]) was significantly higher than that of the control samples (30% [13%–58%], p=0.041). The CSR of the 26 abnormal semen samples that were supplemented with Myo-Ins (38% [20%–50%]) was significantly higher than that of the control samples (23% [12%–30%], p=0.031).ConclusionIn vitro Myo-Ins supplementation of ejaculated human sperm from infertile men resulted in a significant increase in the CSR in samples with abnormal pre-freeze sperm parameters.

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Process and Pitfalls of Sperm Cryopreservation

Cited by 41 publications

References 108 publications

The influence of Percoll® density gradient centrifugation before cryopreservation on the quality of frozen wisent (Bison bonasus) epididymal spermatozoa

The influence of Percoll® density gradient centrifugation before cryopreservation on the quality of frozen wisent (Bison bonasus) epididymal spermatozoa

Cryopreservation Differentially Alters the Proteome of Epididymal and Ejaculated Pig Spermatozoa

Increased cryo-survival rate in ejaculated human sperm from infertile men following pre-freeze in vitro myo-inositol supplementation

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