OBJECTIVE: In vitro cell culture models are widely used in inflammation research; however, information regarding the time- and dose-dependency of inflammatory responses toward LPS in these cell lines is scattered in the literature. MATERIAL: J774A.1 mouse macrophage and THP-1 human monocyte cell lines. TREATMENT: J774A.1 and THP-1 cells were treated with 0-500 ng/mL lipopolysaccharide for 0-24 h. METHODS: SRB and BCA tests were used to measure total protein. Real-time PCR was used to determine gene expression levels, and ELISA was used to assess the protein levels. One-way ANOVA and Tukey's Honestly Significant Difference test were used to test the significance levels. RESULTS: In J774A.1 and THP-1 cells, cytokines responded in distinct patterns upon LPS stimulation in a time- and dose-dependent manner, and the differential regulation of the response to LPS between J774A.1 and THP-1 cells appeared to correlate with the differential regulation of TLR4 at the mRNA level. CONCLUSION: In summary, this study indicated that temporal and dose-dependent responses to LPS need to be controlled for and that extrapolation of data on mechanisms may differ between cell lines of different origin.
OBJECTIVE: In vitro cell culture models are widely used in inflammation research; however, information regarding the time- and dose-dependency of inflammatory responses toward LPS in these cell lines is scattered in the literature. MATERIAL: J774A.1 mouse macrophage and THP-1human monocyte cell lines. TREATMENT: J774A.1 and THP-1 cells were treated with 0-500 ng/mL lipopolysaccharide for 0-24 h. METHODS:SRB and BCA tests were used to measure total protein. Real-time PCR was used to determine gene expression levels, and ELISA was used to assess the protein levels. One-way ANOVA and Tukey's Honestly Significant Difference test were used to test the significance levels. RESULTS: In J774A.1 and THP-1 cells, cytokines responded in distinct patterns upon LPS stimulation in a time- and dose-dependent manner, and the differential regulation of the response to LPS between J774A.1 and THP-1 cells appeared to correlate with the differential regulation of TLR4 at the mRNA level. CONCLUSION: In summary, this study indicated that temporal and dose-dependent responses to LPS need to be controlled for and that extrapolation of data on mechanisms may differ between cell lines of different origin.
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