Protective Role of Alpha-Lipoic Acid During Freezing on the Expression of Apoptosis-Related Genes in Mouse Spermatogonial Stem Cells

Document Type : Original Article

Authors
Kazam Norouzi 1
Hassan Nazari 2
Ebrahim Ahmadi 3
Naser Shams Esfandabadi 4
Azita Afzali 5
1 Faculty of Veterinary Medicine, Shahrekord University
2 Research Institute of Animal Embryo Technology
3 Research Institute of Animal Embryo Technology, Shahrekord university
4 Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahrekord University, Shahrekord- Iran.
5 Department of Anatomical Sciences, Faculty of Medicine, Semnan University of Medical Sciences, Semnan- Iran.
10.22034/ijvcs.2026.15030.1100
Abstract
This study aimed to examine the protective role of alpha-lipoic acid (ALA) against damage caused by freeze-thawing in immature mouse spermatogonial stem cells (SSCs) by analyzing the expression of genes related to apoptosis. SSCs were extracted from the testes of 4–8-day-old C57 mice and, after verification through immunocytochemistry using the PLZF marker and an alkaline phosphatase activity test, were cultured for three weeks. The cells were then frozen in four groups: a control group without antioxidants and three groups treated with different ALA concentrations (0.2, 1, and 5 µM). Following thawing, the expression levels of BAX, BCL2, Caspase-3, and Caspase-9 were measured using Real-time PCR. Findings revealed that 1 µM ALA significantly lowered the expression of the pro-apoptotic genes BAX, Caspase-3, and Caspase-9, while significantly increasing the anti-apoptotic gene BCL2 (P<0.05). The 0.2 µM dose also notably reduced BAX and Caspase-3 expression. However, the highest concentration (5 µM) showed no significant impact. The study concludes that alpha-lipoic acid, especially at 1 µM, can safeguard mouse spermatogonial stem cells from freeze-thaw damage by regulating key apoptosis-related gene expression and may serve as a beneficial additive in the cryopreservation medium for these cells.
Keywords
Spermatogonial stem cells
Alpha-lipoic acid
Cryopreservation
Apoptosis
Gene expression
Mouse
Subjects
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Iranian Journal of Veterinary Clinical Sciences
Volume 19, Issue 2 - Serial Number 33
Fall and Winter 2025/2026
March 2026
Pages 47-60