Prevalidation of Rat GM-CFU Assay for In Vitro Toxicology Application

In vitro haematotoxicity assays are thought to have the potential to significantly reduce and refine the use of animals for haematotoxicity testing. These assays are successful in all types of studies except human toxicology studies in the preclinical setting in which are not required for regulatory testing. Furthermore, these assays could play a key role in bridging the gap between preclinical toxicology studies in animal models and clinical investigations. In previous studies, the Colony Forming Unit-Granulocyte Macrophage ¿CFU-GM¿ assay has been validated for testing drug haematotoxicity (both in mouse bone marrow and human cord blood) and for predicting in vivo human maximal tolerated dose (MTD) by adjusting in vivo data on mouse toxicity. Recently, a Colony Forming Unit-Megakaryocyte (CFU-Mk) assay in human progenitors has also been prevalidated for testing the drug toxicity on megakaryocytes. The rat CFU-GM assay has been used by many researchers for its ability to evaluate in vitro haematotoxicity. Although it has not been developed, a standardised procedure for data comparison could be very important since the rat is the most widely used species for in vivo testing of toxicants. This report presents the results of the prevalidation study developed to analyze the intra-laboratory 15 and inter-laboratories variability of a standardized operating procedure (SOP) for this assay and its performance for the in vitro determination of inhibitory concentration (IC) values of drugs on rat myeloid progenitors (CFU-GM). The results demonstrate that the CFU-GM assay can be applied by using cryopreserved rat bone marrow cells (r BMC) and represents a useful tool for evaluating the toxicity of a compound in terms of both relative toxicity when different molecules are compared and prediction of the degree of in vivo toxicity. Using this assay could greatly reduce the number of rats used in experimental conditions and could also contribute to the accumulation of more toxicity data on compounds to be registered according to the criteria established by the European Registration, Evaluation, Authorization and Restriction of Chemicals (REACH) program.

PESSINA Augusto;  BONOMI Arianna;  CAVICCHINI Loredana;  ALBELLA Beatrix;  CERRATO Laura;  PARENT-MASSIN Dominique;  SIBIRILL Yann;  PARCHMENT Ralph;  BEHRSING Holger;  VERDERIO Paolo;  PIZZAMIGLIO Sara;  GIANGRECO Manuela;  BAGLIO Carolina;  COCCÈ Valentina;  SISTO Francesca;  GRIBALDO Laura; 

2010-05-31

FRAME

JRC56820

0261-1929,   

https://publications.jrc.ec.europa.eu/repository/handle/JRC56820,