ArticleJournal of assisted reproduction and genetics2026
Apoptotic inhibitor Y-27632 improves post-thaw viability and stemness of sheep spermatogonial stem cells.
Article in Journal of assisted reproduction and genetics, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
purposeSpermatogonial stem cells (SSC) are of high significance in animal reproduction, breeding, and regenerative medicine. Cryopreservation of putative SSC is a prerequisite for long-term storage and future applications. However, dissociated putative SSC are very susceptible to cryostress and undergo apoptosis, thereby reducing their functional competence. To minimize the dissociation-induced apoptosis during cryopreservation and post-thaw culture, the role of anti-apoptotic molecule, Y-27632, was evaluated in sheep putative SSC.
methodsThe putative SSC were isolated, enriched, and cultured from prepubertal sheep testis. Y-27632 (0, 5,10, and 15 µM) was added in putative SSC cryomedia (Cy group) and in both freezing and post-thaw culture media (Cy + y group). The effects were compared with non-treated control (C group). One month after cryopreservation, the putative SSC were assessed for viability, ROS production, metabolic activity, stemness associated undifferentiated status, colony characteristics, and apoptosis associated pathway/genes.
resultsThe viability of the dissociated post-thaw putative SSC was significantly higher > 90% when cryopreserved in presence of Y-27632, compared to control (C; 82.5 ± 1.7%). The metabolic activity and stemness associated undifferentiated status were also significantly (p < 0.05) improved in 10 and 15 Cy + y as compared to C. ROCK1 and p53 expression was significantly (p < 0.05) downregulated in 10 Cy + y compared to the C. Also, a significantly (p < 0.01) higher apoptotic cells were observed in C in comparison to 10 Cy and 10 Cy + y (3.9 ± 0.8% vs 1.9 ± 0.2% and 0.35 ± 0.05%).
conclusionThese findings suggest that 10 µM Y-27632, when added to cryomedia and post-thaw culture, reduces apoptosis and thereby improves viability of cells during cryopreservation. It also helps in maintaining undifferentiated status and metabolic activity of sheep putative SSC.
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