基于巨噬细胞实时分析的内毒素检测评价技术研究.PDFVIP

Drug Evaluation Research 第36 卷 第4 期 优先出版 基于巨噬细胞实时分析的内毒素检测评价技术研究 * 赵婧华，庄朋伟，张艳军 ，郝存江 天津中医药大学，天津 300073 摘 要：目的 建立一种基于体外巨噬细胞实时分析的快速、敏感的热原检测技术，为内毒素，即脂多糖 （LPS ）的检测提 供新的方法。方法 体外培养人源性巨噬细胞 THP-1，利用倒置相差显微镜观察 LPS 对巨噬细胞形态变化的影响，然后采 用实时细胞分析仪 （RTCA ）实时监测不同质量浓度的LPS 对巨噬细胞形态变化的影响。结果 LPS 质量浓度≥ 10 pg/mL 时， 可以引起细胞形态变化，实时细胞分析系统可以通过细胞指数（CI 值）实时反映这种形态变化，并且随着剂量的变化，CI 值也呈剂量相关性变化。结论 实时细胞分析系统可以实时监测LPS 对巨噬细胞的影响，为体外检测LPS 提供了一个新的 方法。 关键词：内毒素；实时细胞分析；巨噬细胞；形态变化 中图分类号：R 文献标志码：A 文章编号：1674 - 6376 (20 13) 04 - 0 - 04 DOI: 10.7501/j.issn.1674-6376.2013.04. Measurement of endotoxin through real-time monitoring morphological changes of macrophages ZHAO Jing-hua, ZHUANG Peng-wei, ZHANG Yan-jun, HAO Cun-jiang Tianjin university of traditional Chinese medicine, Tianjin 300073, China Abstract: Objective This article is to establish a rapid, sensitive pyrogen detecting technology based on real-time analysis of macrophages. It provides a new method for detecting endotoxin, that is lipopolysaccharide (LPS). Methods Culture human-derived macrophages (THP-1) in vitro, and observe the impact of LPS on morphological changes of macrophages by using inverted phage contrast microscope. Then monitor the impact of different mass concentrations of LPS on morphological changes of macrophages using real-time cell analyzer (RTCA). Results When the mass concentration of LPS is greater than or equal to 10 pg/mL, LPS can cause cell morphology changes. Real-time analysis system can reflect this morphological changes through CI value, and with the changes of the dose, the CI value also shows a dose-dependent changes. Conclusion Real-time cell analysis system can monitor the impact of LPS on macrophages every time, so a new method of detecting LPS in vitro is provided. Key words: endotoxin; real-time cell analysis; macrophages; morphological changes 细菌