Toward Achieving Harmonization in a Nano-cytotoxicity Assay Measurement through an Interlaboratory Comparison Study
John T. Elliott, Matthias Roesslein, Nam W. Song, Blaza Toman, Agnieszka Kinsner-Ovaskainen, Rawiwan Maniratanachote, Marc L. Salit, Elijah J. Petersen, Fatima Sequeira, Jieun Lee, Francois Rossi, Cordula Hirsch, Harald Krug, Wongsakorn Suchaoin, Peter Wick
Design and development of reliable cell-based nanotoxicology assays is important for ranking of potential hazardous engineered nanomaterials. Challenges to producing a reliable assay protocol include working with nanoparticle dispersions and living cell lines, and the potential for nano- related interference effects. We demonstrate the use of a 96-well plate design for a nano- cytotoxicity MTS cell viability assay. A detailed protocol and an inter-laboratory comparison are used to illustrate the variability of the assay with NH2-polystyrene nanoparticles. Data on both the within and between laboratory system control results can be used to understand the largest sources of variability. This study suggests that a high level of agreement between each of the laboratories can be achieved, but some protocol details such as cell line ID, cultivation, cell rinsing, and nanoparticle dispersion should be considered to ensure comparability of cytotoxicity assays results.