[關(guān)鍵詞]
[摘要]
目的 建立一種基于體外巨噬細(xì)胞實(shí)時(shí)分析的快速、敏感的熱原檢測(cè)技術(shù),為內(nèi)毒素,即脂多糖(LPS)的檢測(cè)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(duì)巨噬細(xì)胞形態(tài)變化的影響,然后采用實(shí)時(shí)細(xì)胞分析儀(RTCA)實(shí)時(shí)監(jiān)測(cè)不同質(zhì)量濃度的LPS對(duì)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(shí),可以引起細(xì)胞形態(tài)變化,實(shí)時(shí)細(xì)胞分析系統(tǒng)可以通過細(xì)胞指數(shù)(CI)值實(shí)時(shí)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(shí)時(shí)細(xì)胞分析系統(tǒng)可以實(shí)時(shí)監(jiān)測(cè)LPS對(duì)巨噬細(xì)胞的影響,從而提供了一個(gè)新的體外檢測(cè)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.
[中圖分類號(hào)]
[基金項(xiàng)目]
重大新藥創(chuàng)制項(xiàng)目(2010ZX09102)