Wang Juan, Zhou Yue, Zhuo Lifan, Yan Bei, Ma Lianghong
Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education, Ningxia Human Sperm Bank, Institute of Medical Sciences, Ningxia Medical University, General Hospital of Ningxia Medical University, Yinchuan, 750004, China.
Department of Urology, General Hospital of Ningxia Medical University, Yinchuan, 750001, China.
Sci Rep. 2025 Jul 2;15(1):22960. doi: 10.1038/s41598-025-00701-3.
The maintenance of DNA integrity in spermatogonial stem cells (SSCs) is essential for ensuring genetic stability and proper function in male germline cells. Here, we established a non-invasive approach to evaluate SSC DNA integrity and investigated the impact of detrimental environmental stressors on genomic stability. SSCs were isolated and cultured from mouse testes. SSCs identity was confirmed through immunofluorescence staining and flow cytometric analysis of specific molecular markers.DNA strand breaks were detected using a novel TdT enzyme-Endo IV-fluorescent probe biosensor, confirming the methodological feasibility. Heat stress and cryopreservation models were established, with the biosensor quantifying stress-induced DNA damage in SSCs. Additionally, extracellular free DNA fragments in culture supernatants were quantified as a non-invasive measure of SSCs DNA integrity.
精原干细胞（SSCs）中DNA完整性的维持对于确保雄性生殖细胞的遗传稳定性和正常功能至关重要。在此，我们建立了一种非侵入性方法来评估SSCs的DNA完整性，并研究有害环境应激源对基因组稳定性的影响。从小鼠睾丸中分离并培养SSCs。通过免疫荧光染色和特定分子标记的流式细胞术分析确认了SSCs的身份。使用新型TdT酶 - 内切酶IV - 荧光探针生物传感器检测DNA链断裂，证实了该方法的可行性。建立了热应激和冷冻保存模型，生物传感器可量化应激诱导的SSCs中的DNA损伤。此外，对培养上清液中的细胞外游离DNA片段进行定量，作为SSCs DNA完整性的非侵入性测量指标。
