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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.
Assuntos

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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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