[关键词]
[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,从而提供了一个新的体外检测LPS的方法。
[Key word]
[Abstract]
Objective To establish a rapid and sensitive pyrogen detecting technology based on real-time analysis of macrophages and provide a new method for detecting endotoxin, i.e. lipopolysaccharide (LPS). Methods Human-derived macrophages THP-1 were cultivated in vitro , and the effect of LPS on morphological changes of macrophages by using inverted phage contrast microscope was observed. Then the impact of LPS at different concentration on the morphological changes of macrophages using real-time cell analyzer (RTCA) was monitored. When the concentration of LPS was greater than or equal to 10 pg/mL, LPS could cause cell morphology changes. Real-time cell analysis system could reflect the morphological changes through cell index (CI) value, and with the changes of the dose, the CI value also showed a dose-dependent change. Conclusion Real-time cell analysis system could monitor the impact of LPS on macrophages, so a new method of detecting LPS in vitro is provided.
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[基金项目]
重大新药创制项目(2010ZX09102)