[關(guān)鍵詞]
[摘要]
目的 建立一種基于體外巨噬細(xì)胞實時分析的快速、敏感的熱原檢測技術(shù),為內(nèi)毒素,即脂多糖(LPS)的檢測提供新的方法。方法 體外培養(yǎng)人源性巨噬細(xì)胞THP-1,利用倒置相差顯微鏡觀察LPS對巨噬細(xì)胞形態(tài)變化的影響,然后采用實時細(xì)胞分析儀(RTCA)實時監(jiān)測不同質(zhì)量濃度的LPS對巨噬細(xì)胞形態(tài)變化的影響。結(jié)果 LPS質(zhì)量濃度≥10 pg/mL時,可以引起細(xì)胞形態(tài)變化,實時細(xì)胞分析系統(tǒng)可以通過細(xì)胞指數(shù)(CI)值實時反映這種形態(tài)變化,并且隨著劑量的變化,CI值也呈劑量相關(guān)性變化。結(jié)論 實時細(xì)胞分析系統(tǒng)可以實時監(jiān)測LPS對巨噬細(xì)胞的影響,從而提供了一個新的體外檢測LPS的方法。
[Key word]
[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.
[中圖分類號]
[基金項目]
重大新藥創(chuàng)制項目(2010ZX09102)