The effect of cryopreservation on DNA methylation patterns of the chromosome 15q11-q13 region in human spermatozoa.

Abstract:

:Human sperm cryopreservation is a common technique which is used in assisted reproductive technologies. Despite the existence of evidence supporting the production of ROS and DNA fragmentation during sperm cryopreservation, there is little and equivocal information about the cryopreservation effects on methylation of imprinted genes and imprinting control regions. In this study, we have investigated the effects of cryopreservation on DNA methylation in promoter regions of SNURF-SNRPN and UBE3A imprinted genes, PWS-ICR and AS-ICR in the chromosome 15q11-q13 region. Semen samples from 10 healthy normozoospermic men were collected and each sample was divided into four equal aliquots: fresh, cryoprotectant, cryopreservation, and H2O2. We measured the ROS levels and DNA fragmentation using DCFH-DA and TUNEL assay respectively by flow cytometry. DNA methylation in promoter regions of SNURF-SNRPN and UBE3A imprinted genes, PWS-ICR and AS-ICR in the chromosome 15q11-q13 region were evaluated by quantitative methylation-specific PCR technique. Intracellular levels of ROS and percentage of TUNEL-positive spermatozoa significantly increased in cryopreservation group compared to fresh group. Exposure to cryoprotectant had no significant effect on ROS levels and DNA fragmentation. Neither cryopreservation nor exposure to cryoprotectant significantly affected DNA methylation of the selected gene regions. However, DNA fragmentation had positive correlation with DNA methylation of AS-ICR. In conclusion, based on our study, clinical use of sperm cryopreservation for fertility treatments appear to be safe in regard to DNA methylation in the chromosome 15q11-q13 region.

journal_name

Cell Tissue Bank

journal_title

Cell and tissue banking

authors

Khosravizadeh Z,Hassanzadeh G,Tavakkoly Bazzaz J,Alizadeh F,Totonchi M,Salehi E,Khodamoradi K,Khanehzad M,Hosseini SR,Abolhassani F

doi

10.1007/s10561-020-09828-1

subject

Has Abstract

pub_date

2020-09-01 00:00:00

pages

433-445

issue

3

eissn

1389-9333

issn

1573-6814

pii

10.1007/s10561-020-09828-1

journal_volume

21

pub_type

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