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模拟空间相关环境下神经胶质细胞外泌体的可视化快速检测

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目的 探讨模拟空间环境下神经胶质细胞外泌体的可视化快速检测方法.方法 通过2 Gy、5 Gy辐照,12h、24 h微重力的处理方式,建立模拟空间环境下神经胶质细胞的损伤模型,将条件培养基和试剂盒提取的外泌体转移至神经元,明确神经胶质细胞对神经元产生的影响.通过膜嵌入型荧光染料分子对外泌体进行活细胞荧光标记,以荧光强度为指标对条件培养基中的外泌体释放规律进行可视化监测.探究不同保存条件、保存时间对外泌体的数量和粒径的影响.结果 首先,本实验证实模拟空间环境下受损胶质细胞释放的胞外囊泡可在一定程度上保护神经元,而外泌体在这一过程中发挥决定性作用,并确定神经系统外泌体可视化检测方案与传统外泌体验证方案的结论一致;其次,本实验建立了可用于半定量分析的外泌体数量和粒径经验曲线,为细胞培养基中外泌体的快速检测提供了新思路.最后,本实验明确了培养基样本的最佳保存条件,为航天飞行后培养基样本的地基/天基在线分析奠定了基础.结论 通过荧光标记可实现外泌体的快速检测和分析,并用于模拟空间环境下神经胶质细胞损伤的探究.
Visualized and rapid detection of extracellular vesicles of glial cells in a simulated space environment
Objective To rapidly visualize and detect the extracellular vesicles of glial cells in a simulated space environment.Methods By using 2,5 Gy irradiation and 12,24 h microgravity treatment,a damage model of glial cells was established in a simulated space environment.Exosomes extracted from conditioned media with reagent kits were transferred to neurons to elucidate the impact of glial cells on neurons.By performing live-cell fluorescence labeling of exosomes,a visualization monitoring scheme based on fluorescence intensity analysis was developed to characterize the release patterns of extracellular vesicles.The release patterns of exosomes were represented by the fluorescence intensity of the conditioned media.The effects of different storage conditions and duration on the quantity and size of exosomes were investigated.Results The exosomes released from damaged glial cells in the simulated space environment could to some extent protect neurons,with exosomes playing a decisive role in this process,and the neurosystem exosome visualization detection scheme was consistent with the traditional exosome validation scheme.An empirical curve for exosome quantity and size was established for semi-quantitative analysis,providing a new approach for rapid detection of exosomes in cell culture media.Furthermore,the optimal storage conditions for culture medium samples were clarified,laying the foundation for ground/space-based online analysis of culture medium samples after spaceflight.Conclusion Exosome rapid detection and analysis can be achieved through fluorescence labeling and utilized to investigate glial cell injury in a simulated space environment.

space environmentneuroglial cellextracellular vesiclefluorescent probevisualized detection

尔天漪、蓝钰、刘倍沁、王舒钥、赵亚丽、杨成佳、马宏

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北京理工大学医学技术学院,北京 100081

中国航天员科研训练中心,北京 100094

空间环境 神经胶质细胞 外泌体 荧光探针 可视化检测

载人航天工程航天医学实验领域项目

HYZHXM02003

2024

航天医学与医学工程
中国航天员科研训练中心

航天医学与医学工程

影响因子:0.392
ISSN:1002-0837
年,卷(期):2024.35(1)
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