In Vitro Bioassay of Endotoxin Using Fluorescein as a pH Indicator in a Macrophage Cell Culture System
Yonsei Medical Journal
;
: 268-274, 2005.
Artigo
em Inglês
| 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.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Bioensaio
/
Estudo Comparativo
/
Células Cultivadas
/
Lipopolissacarídeos
/
Meios de Contraste
/
Fluoresceína
/
Meios de Cultura
/
Endotoxinas
/
Concentração de Íons de Hidrogênio
/
Teste do Limulus
Limite:
Animais
Idioma:
Inglês
Revista:
Yonsei Medical Journal
Ano de publicação:
2005
Tipo de documento:
Artigo
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