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Yonsei medical journal v.46 no.2, 2005년, pp.268 - 274  

In Vitro Bioassay of Endotoxin Using Fluorescein as a pH Indicator in a Macrophage Cell Culture System

Lee, Dong Hee    (Department of Medical Engineering, Yonsei University College of Medicine, Seoul, Korea.   ); Sung, Hak-Joon    (Department of Medical Engineering, Yonsei University College of Medicine, Seoul, Korea.   ); Han, Dong-Wook    (Department of Medical Engineering, Yonsei University College of Medicine, Seoul, Korea.   ); Lee, Min-Sub    (Department of Medical Engineering, Yonsei University College of Medicine, Seoul, Korea.   ); Ryu, Gyu Ha    (Department of Medical Devices & Radiation Health, Korea Food & Drug Administration, Seoul, Korea.   ); Aihara, Maki    (Division of Microbiology, National Institute of Health Sciences, Tokyo, Japan.   ); Takatori, Kosuke    (Division of Microbiology, National Institute of Health Sciences, Tokyo, Japan.   ); Park, Jong-Chul    (Department of Medical Engineering, Yonsei University College of Medicine, Seoul, Korea.  );
  • 초록

    Based on the biological activity of endotoxin, we propose a possible new method for detecting endotoxin using a pH-indication system of macrophage culture media. After RAW 264.7 macrophage cells were treated with lipopolysaccharide (LPS), the addition of fluorescein to the LPS-treated media reproductively reduced its absorption and emission spectra (it was a dose-dependent reduction). The advantages of this LPS-detection method were compared with the Limulus Amebocyte Lysate (LAL) test by using purified bacterial LPS ( Salmonella minnessota , Escherichia coli , and Pseudomonas aeruginosa ). Additionally, the absorption and fluorescence intensity of fluorescein, following treatment of RAW 264.7 cells with a high concentration of Staphylococcus aureus (Gram-positive, lysed bacteria), could not generally be detected by the LAL test, but they were found to be reduced, in a dose-response relationship, with this new system. The macrophage culture system-method might be a good supplement to the LAL assay for detection of LPS, Gram-negative and Gram-positive bacteria.


  • 주제어

    Lipopolysaccharide .   macrophage .   fluorescein .   Limulus Amebocyte Lysate test .   Staphylococcus aureus .   pH.  

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