In Vitro Bioassay of Endotoxin Using Fluorescein as a pH Indicator in a Macrophage Cell Culture System
Yonsei Medical Journal
;
: 268-274, 2005.
Article
Dans Anglais
| WPRIM
| ID: wpr-99091
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 (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.
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Dosage biologique
/
Étude comparative
/
Cellules cultivées
/
Lipopolysaccharides
/
Produits de contraste
/
Fluorescéine
/
Milieux de culture
/
Endotoxines
/
Concentration en ions d'hydrogène
/
Test LAL
Limites du sujet:
Animaux
langue:
Anglais
Texte intégral:
Yonsei Medical Journal
Année:
2005
Type:
Article
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