Evaluation of Nrf2/ARE Signaling Pathway in the Presence of Pentoxifylline as a Cryoprotectant in Mouse Spermatogonial Stem Cells

• DOI: 10.1089/bio.2021.0167
• Publication Date: 2022-08-25T15:37:44Z
• Authors: Ghazaleh Sadeghiani; Maryam Khanehzad; Mohammad Ali Sadi...; Fardin Amidi; Mehrnoush Malekzadeh; Tayebeh Rastegar
• Keywords: Cryoprotectant; Viability assay; Malondialdehyde

The process of spermatogonial stem cell cryopreservation (SSCs) in young male cancer survivors is associated with increased reactive oxygen species (ROS), DNA fragmentation, apoptosis, decreased activity, and finally reduced fertility SSCs. Therefore, it necessary to add cryoprotectants the freezing medium minimize injuries cryopreservation. In addition, Nrf2/ARE pathway a main cellular that regulates antioxidant defense system. purpose this study was evaluate cryoprotective effect pentoxifylline (PTX) on SSCs after freezing-thawing through pathway. extracted from neonatal mice testes were isolated their purity measured by flow cytometry GDNF family receptor alpha-1 (GFRα1) inhibitor differentiation 4 (ID4). After culturing, cells frozen different groups for 1 month. freezing-thawing, viability, colonization rate, intracellular ROS, malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) evaluated. Quantitative real-time polymerase chain reaction western blotting done assess expression levels Nrf2, Keap-1, PI3K, AKT genes proteins. survival rates SSCs, SOD, CAT levels, significantly higher PTX group compared other groups. Keap-1 level ROS MDA production also (p-value <0.05). According our findings, can activate signaling pathway; therefore, could be suitable cryoprotectant candidate long-term storage clinical setting.