Modern human sperm freezing: Effect on DNA, chromatin and acrosome integrity

Objective: Presence of vitrification method in sperm freezing and the introduction of solid surface vitrification beside rapid freezing in vapour, opens an easy and safe way to help infertility centres. While the effects of cryopreservation on motility, morphology and viability of sperm are document...

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Main Authors: Tahereh Rahiminia, Akram Hosseini, Morteza Anvari, Saeed Ghasemi-Esmailabad, Ali Reza Talebi
Format: Article
Language:English
Published: Elsevier 2017-08-01
Series:Taiwanese Journal of Obstetrics & Gynecology
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S102845591730133X
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author Tahereh Rahiminia
Akram Hosseini
Morteza Anvari
Saeed Ghasemi-Esmailabad
Ali Reza Talebi
author_facet Tahereh Rahiminia
Akram Hosseini
Morteza Anvari
Saeed Ghasemi-Esmailabad
Ali Reza Talebi
author_sort Tahereh Rahiminia
collection DOAJ
description Objective: Presence of vitrification method in sperm freezing and the introduction of solid surface vitrification beside rapid freezing in vapour, opens an easy and safe way to help infertility centres. While the effects of cryopreservation on motility, morphology and viability of sperm are documented, the question of the probable alteration of sperm DNA, chromatin and acrosome integrity after freezing and thawing procedures in different methods is still controversial. Materials and methods: Normal sample were collected according to WHO strict criteria. Sperm suspensions were mixed 1:1 with 0.5 M sucrose and divided into four equal aliquots for freezing: fresh, nitrogen direct immersion vitrification (Vit), solid surface vitrification (SSV) and in vapour (Vapour). Sperm suspensions were transferred into a 0.25 ml sterile plastic. Then straw was inserted inside the 0.5 ml straw. For thawing, the straws were immersed in a 42 °C water bath. Beside the sperm parameters, we assessed the acrosome reaction by double staining, chromatin integrity by toluidine blue (Tb) and chromomycin A3 (CMA3) and DNA integrity by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) respectively. Results: In progressive motility, the highest rate occurred in Vit (39.9 ± 13.3). Moreover, the lowest rate of immotile sperm was in Vit (32.7 ± 16.3). In normal morphology, the group Vit was similar to the fresh, while SSV and Vapour were significantly different from the fresh. The percentage of acrosome-reacted sperms was more in Vit (81.3 ± 10.2) than the fresh group. TUNEL+ results showed that DNA fragmentation was significantly increased in Vit (p-value = 0.025). While in SSV and Vapour results were comparable to fresh. There was a significant correlation between TUNEL+ and normal morphology, TB, CMA3 and presence of intact acrosome. Conclusion: Sperm in Vapour was healthier in terms of DNA, chromatin and acrosome integrity. In contrast of higher motility and normal morphology; DNA, chromatin and acrosome integrity were decreased in Vit. However, these findings were more acceptable in SSV or Vapour.
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spelling doaj.art-9161fc5686624f9daa079e7f16d1d7e52022-12-22T00:00:47ZengElsevierTaiwanese Journal of Obstetrics & Gynecology1028-45592017-08-0156447247610.1016/j.tjog.2017.02.004Modern human sperm freezing: Effect on DNA, chromatin and acrosome integrityTahereh RahiminiaAkram HosseiniMorteza AnvariSaeed Ghasemi-EsmailabadAli Reza TalebiObjective: Presence of vitrification method in sperm freezing and the introduction of solid surface vitrification beside rapid freezing in vapour, opens an easy and safe way to help infertility centres. While the effects of cryopreservation on motility, morphology and viability of sperm are documented, the question of the probable alteration of sperm DNA, chromatin and acrosome integrity after freezing and thawing procedures in different methods is still controversial. Materials and methods: Normal sample were collected according to WHO strict criteria. Sperm suspensions were mixed 1:1 with 0.5 M sucrose and divided into four equal aliquots for freezing: fresh, nitrogen direct immersion vitrification (Vit), solid surface vitrification (SSV) and in vapour (Vapour). Sperm suspensions were transferred into a 0.25 ml sterile plastic. Then straw was inserted inside the 0.5 ml straw. For thawing, the straws were immersed in a 42 °C water bath. Beside the sperm parameters, we assessed the acrosome reaction by double staining, chromatin integrity by toluidine blue (Tb) and chromomycin A3 (CMA3) and DNA integrity by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) respectively. Results: In progressive motility, the highest rate occurred in Vit (39.9 ± 13.3). Moreover, the lowest rate of immotile sperm was in Vit (32.7 ± 16.3). In normal morphology, the group Vit was similar to the fresh, while SSV and Vapour were significantly different from the fresh. The percentage of acrosome-reacted sperms was more in Vit (81.3 ± 10.2) than the fresh group. TUNEL+ results showed that DNA fragmentation was significantly increased in Vit (p-value = 0.025). While in SSV and Vapour results were comparable to fresh. There was a significant correlation between TUNEL+ and normal morphology, TB, CMA3 and presence of intact acrosome. Conclusion: Sperm in Vapour was healthier in terms of DNA, chromatin and acrosome integrity. In contrast of higher motility and normal morphology; DNA, chromatin and acrosome integrity were decreased in Vit. However, these findings were more acceptable in SSV or Vapour.http://www.sciencedirect.com/science/article/pii/S102845591730133XVitrificationSolid surface vitrificationDNA integrityChromatin integrityAcrosome integrity
spellingShingle Tahereh Rahiminia
Akram Hosseini
Morteza Anvari
Saeed Ghasemi-Esmailabad
Ali Reza Talebi
Modern human sperm freezing: Effect on DNA, chromatin and acrosome integrity
Taiwanese Journal of Obstetrics & Gynecology
Vitrification
Solid surface vitrification
DNA integrity
Chromatin integrity
Acrosome integrity
title Modern human sperm freezing: Effect on DNA, chromatin and acrosome integrity
title_full Modern human sperm freezing: Effect on DNA, chromatin and acrosome integrity
title_fullStr Modern human sperm freezing: Effect on DNA, chromatin and acrosome integrity
title_full_unstemmed Modern human sperm freezing: Effect on DNA, chromatin and acrosome integrity
title_short Modern human sperm freezing: Effect on DNA, chromatin and acrosome integrity
title_sort modern human sperm freezing effect on dna chromatin and acrosome integrity
topic Vitrification
Solid surface vitrification
DNA integrity
Chromatin integrity
Acrosome integrity
url http://www.sciencedirect.com/science/article/pii/S102845591730133X
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