In vitro bioassay of endotoxin using fluorescein as a pH indicator in a macrophage cell culture system

Dong Hee Lee, Hak Joon Sung, Dong Wook Han, Min Sub Lee, Gyu Ha Ryu, Maki Aihara, Kosuke Takatori, Jong Chul Park

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

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 (Grain-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.

Original languageEnglish
Pages (from-to)268-274
Number of pages7
JournalYonsei medical journal
Volume46
Issue number2
DOIs
Publication statusPublished - 2005 Apr 30

All Science Journal Classification (ASJC) codes

  • Medicine(all)

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