Non-invasive assessment of stem cell DNA integrity in response to environmental stressors.

IF 3.9 2区 综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES
Juan Wang, Yue Zhou, Lifan Zhuo, Bei Yan, Lianghong Ma
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Abstract

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.

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干细胞DNA完整性对环境应激反应的非侵入性评估。
精原干细胞(ssc) 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。在这里,我们建立了一种非侵入性的方法来评估SSC DNA的完整性,并研究了有害环境应激源对基因组稳定性的影响。从小鼠睾丸中分离培养ssc。通过免疫荧光染色和流式细胞术分析特异性分子标记,确认SSCs的身份。采用新型TdT酶- endo iv荧光探针生物传感器检测DNA链断裂,证实了方法的可行性。建立热应激和低温保存模型,用生物传感器量化应力诱导的ssc DNA损伤。此外,培养上清液中的细胞外游离DNA片段被定量作为SSCs DNA完整性的非侵入性测量。
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来源期刊
Scientific Reports
Scientific Reports Natural Science Disciplines-
CiteScore
7.50
自引率
4.30%
发文量
19567
审稿时长
3.9 months
期刊介绍: We publish original research from all areas of the natural sciences, psychology, medicine and engineering. You can learn more about what we publish by browsing our specific scientific subject areas below or explore Scientific Reports by browsing all articles and collections. Scientific Reports has a 2-year impact factor: 4.380 (2021), and is the 6th most-cited journal in the world, with more than 540,000 citations in 2020 (Clarivate Analytics, 2021). •Engineering Engineering covers all aspects of engineering, technology, and applied science. It plays a crucial role in the development of technologies to address some of the world''s biggest challenges, helping to save lives and improve the way we live. •Physical sciences Physical sciences are those academic disciplines that aim to uncover the underlying laws of nature — often written in the language of mathematics. It is a collective term for areas of study including astronomy, chemistry, materials science and physics. •Earth and environmental sciences Earth and environmental sciences cover all aspects of Earth and planetary science and broadly encompass solid Earth processes, surface and atmospheric dynamics, Earth system history, climate and climate change, marine and freshwater systems, and ecology. It also considers the interactions between humans and these systems. •Biological sciences Biological sciences encompass all the divisions of natural sciences examining various aspects of vital processes. The concept includes anatomy, physiology, cell biology, biochemistry and biophysics, and covers all organisms from microorganisms, animals to plants. •Health sciences The health sciences study health, disease and healthcare. This field of study aims to develop knowledge, interventions and technology for use in healthcare to improve the treatment of patients.
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