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EC number: 214-675-6 | CAS number: 1184-78-7
Table1 Overview Stimulation index of CD86 and Cell Viability in Experiment 1 and 2 of TMAO anhydrous
% Viability (Mean)*
Colour Interference S.I.*
- Not Applicable
Table2 Overview Stimulation index of CD86 and Cell Viability in Experiment 1 and 2 of the Positive, Negative and Vehicle Control
IgG1 value (%)
CD86 basal expression (%)
RPMI Mean Viability
* Red values are either below 70% viability, above 150S.I..
Table3 Overview EC150and CV70Values
Test item Experiment 1
Test item Experiment 2
NA = Not applicable
The objective of this in vitro GLP-compliant study is to evaluate the ability of TMAO anhydrous to increase the expression levels of CD86 cell surface marker in the U937 cell line in the U937 cell line activation Test (U-Sens™) assay.
TMAO anhydrous was a white powder. TMAO anhydrous was dissolved in complete medium at 0.4 mg/mL. In the first experiment the stock was diluted to six test concentrations (1, 10, 20, 50, 100 and 200 μg/mL). In the second experiment, a more narrow dose-response analysis was performed up to 200 µg/mL. Test item, positive control and negative control were exposed to the cells during 45±3 hours. After the incubation with test item or controls, cells were treated with IgG1 CD86 specific antibody. Two independent experiments were performed.
Both experiments passed the acceptance criteria:
• At the end of the incubation treatment period, the mean viability of the triplicate untreated U937 cells was above the threshold of 90% (100% in experiment 1 and 2).
• The CD86 basal expression of untreated U937 cells is within the range of ≥ 2% and ≤ 25% in both experiments.
• At least two out of three IgG1 values of untreated U937 cells fell within the range of ≥ 0.6% and < 1.5% in both experiments.
• No drift in CD86 expression was observed in the untreated controls (RPMI) and negative (LA) controls.
In both experiments the positive (TNBS) and negative (LA) control were considered valid. Overall it is concluded that the test conditions were adequate and that the test system functioned properly.
TMAO anhydrous showed no toxicity (No CV70 value) and no biologically relevant induction of the CD86 activity (No EC150 value) was measured at any of the test concentrations in both experiments. TMAO anhydrous is classified as negative in the U-Sens™ assay since negative results (< 150% increase) were observed at all test concentrations with a cell viability of >70% compared to the vehicle control.
In conclusion, TMAO anhydrous is classified as negative (no increase in the expression levels of CD86 cell surface marker in the U937 cell line) under the described experimental conditions
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