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Development of a Partition-Controlled Dosing System for Cell Assays

Hydrophobic and volatile chemicals have proven to be difficult to dose in cell assays. Cosolvents are often needed to dissolve these chemicals in cell culture medium. Moreover, the free concentration of these chemicals in culture medium may diminish over time due to metabolism, evaporation, and nonspecific binding to well plate surfaces and serum constituents. The aim of this study was to develop a partition-controlled dosing system to maintain constant concentrations of benzo(a)pyrene, 1,2-dichlorobenzene, and 1,2,4-trichlorobenzene in an ethoxyresorufin-O-deethylase (EROD) assay and a cytotoxicity assay with the rainbow trout (Oncorhynchus mykiss) cell lines RTL-W1 and RTgill-W1. Polydimethylsiloxane (PDMS) sheets were loaded with test chemicals in a spiked methanol/water solution and placed in the wells, filled with culture medium, of a 24-well culture plate. Cells were grown on inserts and were subsequently added to the wells with the PDMS sheets. The system reached equilibrium within 24 h, even for t...

Authors:   Nynke I. Kramer; Frans J. M. Busser; Mattheus T. T. Oosterwijk; Kristin Schirmer; Beate I. Escher; Joop L. M. Hermens
Journal:   Chemical Research in Toxicology
Year:   2010
DOI:   10.1021/tx1002595
Publication date:   15-Nov-2010
Facts, background information, dossiers
  • cell lines
  • cell culture
  • cell culturing
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