Validation of a high throughput flow cytometric in vitro micronucleus assay including assessment of metabolic activation in TK6 cells

2014 ◽  
Vol 55 (9) ◽  
pp. 704-718 ◽  
Author(s):  
Annemette V. Thougaard ◽  
Joan Christiansen ◽  
Tomas Mow ◽  
Jorrit J. Hornberg
Keyword(s):  
High Throughput ◽  
Micronucleus Assay ◽  
Metabolic Activation ◽  
Flow Cytometric ◽  
Tk6 Cells ◽  
In Vitro Micronucleus Assay

Flow cytometric 96-well microplate-based in vitro micronucleus assay with human TK6 cells: Protocol optimization and transferability assessment

2013 ◽  
Vol 54 (3) ◽  
pp. 180-194 ◽  
Author(s):  
Steven M. Bryce ◽  
Svetlana L. Avlasevich ◽  
Jeffrey C. Bemis ◽  
Matthew Tate ◽  
Richard M. Walmsley ◽  
...  
Keyword(s):  
Micronucleus Assay ◽  
Flow Cytometric ◽  
Protocol Optimization ◽  
Tk6 Cells ◽  
In Vitro Micronucleus Assay

scholarly journals Analysis of negative historical control group data from the in vitro micronucleus assay using TK6 cells

2018 ◽  
Vol 825 ◽  
pp. 40-50 ◽  
Author(s):  
David P. Lovell ◽  
Mick Fellows ◽  
Francesco Marchetti ◽  
Joan Christiansen ◽  
Azeddine Elhajouji ◽  
...  
Keyword(s):  
Micronucleus Assay ◽  
Historical Control Group ◽  
Historical Control ◽  
Control Group ◽  
Group Data ◽  
Tk6 Cells ◽  
In Vitro Micronucleus Assay

A flow cytometry based in vitro micronucleus assay in TK6 cells-Validation using early stage pharmaceutical development compounds

2010 ◽  
Vol 52 (5) ◽  
pp. 363-372 ◽  
Author(s):  
Magdalena Lukamowicz ◽  
Katherine Woodward ◽  
Micheline Kirsch-Volders ◽  
Willi Suter ◽  
Azeddine Elhajouji
Keyword(s):  
Flow Cytometry ◽  
Early Stage ◽  
Micronucleus Assay ◽  
Pharmaceutical Development ◽  
Tk6 Cells ◽  
In Vitro Micronucleus Assay

scholarly journals Interlaboratory evaluation of a flow cytometric, high content in vitro micronucleus assay

2008 ◽  
Vol 650 (2) ◽  
pp. 181-195 ◽  
Author(s):  
Steven M. Bryce ◽  
Svetlana L. Avlasevich ◽  
Jeffrey C. Bemis ◽  
Magdalena Lukamowicz ◽  
Azeddine Elhajouji ◽  
...  
Keyword(s):  
Micronucleus Assay ◽  
Flow Cytometric ◽  
In Vitro Micronucleus Assay

Development and validation of an in vitro micronucleus assay platform in TK6 cells

2012 ◽  
Vol 746 (1) ◽  
pp. 29-34 ◽  
Author(s):  
Zhanna Sobol ◽  
Michael L. Homiski ◽  
Donna A. Dickinson ◽  
Richard A. Spellman ◽  
Dingzhou Li ◽  
...  
Keyword(s):  
Micronucleus Assay ◽  
Tk6 Cells ◽  
Development And Validation ◽  
In Vitro Micronucleus Assay

Multi-lab validation of an improved flow cytometric in vitro micronucleus assay using attached cell lines

2009 ◽  
Vol 189 ◽  
pp. S84
Author(s):  
Robert Young ◽  
Jing Shi ◽  
Steven Bryce ◽  
John Nicolette ◽  
Marilyn Diehl ◽  
...  
Keyword(s):  
Cell Lines ◽  
Micronucleus Assay ◽  
Flow Cytometric ◽  
Attached Cell ◽  
In Vitro Micronucleus Assay

High Content Analysis of an In Vitro Model for Metabolic Toxicity

2014 ◽  
Vol 19 (10) ◽  
pp. 1402-1408 ◽  
Author(s):  
Stephanie D. Cole ◽  
Janna S. Madren-Whalley ◽  
Albert P. Li ◽  
Russell Dorsey ◽  
Harry Salem
Keyword(s):  
Content Analysis ◽  
High Throughput ◽  
Metabolic Activation ◽  
Hepatic Metabolism ◽  
Cell Types ◽  
Multiple Organ ◽  
Compound Libraries ◽  
High Content Analysis ◽  
Calcein Am

In vitro models that accurately and rapidly assess hepatotoxicity and the effects of hepatic metabolism on nonliver cell types are needed by the U.S. Department of Defense and the pharmaceutical industry to screen compound libraries. Here, we report the first use of high content analysis on the Integrated Discrete Multiple Organ Co-Culture (IdMOC) system, a high-throughput method for such studies. We cultured 3T3-L1 cells in the presence and absence of primary human hepatocytes, and exposed the cultures to 4-aminophenol and cyclophosphamide, model toxicants that are respectively detoxified and activated by the liver. Following staining with calcein-AM, ethidium homodimer-1, and Hoechst 33342, high content analysis of the cultures revealed four cytotoxic endpoints: fluorescence intensities of calcein-AM and ethidium homodimer-1, nuclear area, and cell density. Using these endpoints, we observed that the cytotoxicity of 4-aminophenol in 3T3-L1 cells in co-culture was less than that observed for 3T3-L1 monocultures, consistent with the known detoxification of 4-aminophenol by hepatocytes. Conversely, cyclophosphamide cytotoxicity for 3T3-L1 cells was enhanced by co-culturing with hepatocytes, consistent with the known metabolic activation of this toxicant. The use of IdMOC plates combined with high content analysis is therefore a multi-endpoint, high-throughput capability for measuring the effects of metabolism on toxicity.

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