@article { author = {Khosronezhad, Nahid and Hassanzadeh, Vahideh and Hezavehei, Maryam and Shahverdi, Abdolhossein and Shahhoseini, Maryam}, title = {Comparative Epigenetic Analysis of Imprinting Genes Involved in Fertility, in Cryopreserved Human Sperms with Rapid Freezing versus Vitrification Methods}, journal = {Cell Journal (Yakhteh)}, volume = {25}, number = {4}, pages = {238-246}, year = {2023}, publisher = {Royan Institute, Iranian Academic Center for Education Culture and Research (ACECR)}, issn = {2228-5806}, eissn = {2228-5814}, doi = {10.22074/cellj.2023.1974291.1171}, abstract = {Objective: Choosing the optimal method for human sperm cryopreservation seems necessary to reduce cryoinjury.The aim of this study is to compare two cryopreservation methods including rapid-freezing and vitrification, in termsof cellular parameters, epigenetic patterns and expression of paternally imprinted genes (PAX8, PEG3 and RTL1) inhuman sperm which play a role in male fertility.Materials and Methods: In this experimental study, semen samples were collected from 20 normozoospermic men.After washing the sperms, cellular parameters were investigated. DNA methylation and expression of genes wereinvestigated using methylation-specific polymerase chain reaction (PCR) and real-time PCR methods, respectively.Results: The results showed a significant decrease in sperm motility and viability, while a significant increase wasobserved in DNA fragmentation index of cryopreserved groups in comparison with the fresh group. Moreover, a significantreduction in sperm total motility (TM, P<0.01) and viability (P<0.01) was determined, whereas a significant increasewas observed in DNA fragmentation index (P<0.05) of the vitrification group compared to the rapid-freezing group.Our results also showed a significant decrease in expression of PAX8, PEG3 and RTL1 genes in the cryopreservedgroups compared to the fresh group. However, expression of PEG3 (P<0.01) and RTL1 (P<0.05) genes were reducedin the vitrification compared to the rapid-freezing group. Moreover, a significant increase in the percentage of PAX8,PEG3 and RTL1 methylation was detected in the rapid-freezing group (P<0.01, P<0.0001 and P<0.001, respectively)and vitrification group (P<0.01, P<0.0001 and P<0.0001, respectively) compared to the fresh group. Additionally,percentage of PEG3 and RTL1 methylation in the vitrification group was significantly increased (P<0.05 and P<0.05,respectively) compared to the rapid-freezing group.Conclusion: Our findings showed that rapid-freezing is more suitable method for maintaining sperm cell quality. Inaddition, due to the role of these genes in fertility, changes in their expression and epigenetic modification may affectfertility.}, keywords = {Epigenetics,Male Fertility,Rapid-freezing,Vitrification}, url = {https://www.celljournal.org/article_701189.html}, eprint = {https://www.celljournal.org/article_701189_57e3a303c43015581398c0c6b237d25f.pdf} }