Repository of Research and Investigative Information

Repository of Research and Investigative Information

Ilam University of Medical Sciences

Melatonin in cryopreservation media improves transplantation efficiency of frozen-thawed spermatogonial stem cells into testes of azoospermic mice

Thu Feb 2 17:54:51 2023

(2022) Melatonin in cryopreservation media improves transplantation efficiency of frozen-thawed spermatogonial stem cells into testes of azoospermic mice. Stem Cell Research & Therapy. p. 15.

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Abstract

Background Cryostorage of spermatogonial stem cells (SSCs) is an appropriate procedure for long-term storage of SSCs for fertility preservation. However, it causes damage to cellular structures through overproduction of ROS and oxidative stress. In this study, we examined the protective effect of melatonin as a potent antioxidant in the basic freezing medium to establish an optimal cryopreservation method for SSCs. Methods SSCs were obtained from the testes of neonatal male mice aged 3-6 days. Then, 100 mu M melatonin was added to the basic freezing medium containing DMSO for cryopreservation of SSCs. Viability, apoptosis-related markers (BAX and BCL2), and intracellular ROS generation level were measured in frozen-thawed SSCs before transplantation using the MTT assay, immunocytochemistry, and flow cytometry, respectively. In addition, Western blotting and immunofluorescence were used to evaluate the expression of proliferation (PLZF and GFR alpha 1) and differentiation (Stra8 and SCP3) proteins in frozen-thawed SSCs after transplantation into recipient testes. Results The data showed that adding melatonin to the cryopreservation medium markedly increased the viability and reduced intracellular ROS generation and apoptosis (by decreasing BAX and increasing BCL2) in the frozen-thawed SSCs (p < 0.05). The expression levels of proliferation (PLZF and GFR alpha 1) and differentiation (Stra8 and SCP3) proteins and resumption of spermatogenesis from frozen-thawed SSCs followed the same pattern after transplantation. Conclusions The results of this study revealed that adding melatonin as an antioxidant to the cryopreservation medium containing DMSO could be a promising strategy for cryopreservation of SSCs to maintain fertility in prepubertal male children who suffer from cancer.

Item Type: Article
Creators:
CreatorsEmail
Kazemzadeh, S.UNSPECIFIED
Mohammadpour, S.UNSPECIFIED
Madadi, S.UNSPECIFIED
Babakhani, A.UNSPECIFIED
Shabani, M.UNSPECIFIED
Khanehzad, M.UNSPECIFIED
Keywords: Melatonin Spermatogonial stem cells Cryopreservation Transplantation lipid-peroxidation hydrogen-peroxide sperm spermatogenesis fertility survival restoration receptors recovery quality Cell Biology Research & Experimental Medicine
Divisions:
Page Range: p. 15
Journal or Publication Title: Stem Cell Research & Therapy
Journal Index: ISI
Volume: 13
Number: 1
Identification Number: https://doi.org/10.1186/s13287-022-03029-1
Depositing User: مهندس مهدی شریفی
URI: http://eprints.medilam.ac.ir/id/eprint/3966

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