Cytogenetic characterization of the transgenic Big Blue Rat2 and Big Blue mouse embryonic fibroblast cell lines.

Abstract

The transgenic Big Blue Rat2 and Big Blue mouse embryonic fibroblast cell lines have been used to complement the transgenic Big Blue rat and mouse in vivo mutagenesis assays. However, limited information is available regarding the karyology of these cell lines. Therefore, we have characterized the ploidy, mitotic index, spontaneous frequencies of chromosome and chromatid aberrations and rate of micronucleus (MN) formation in both cell lines. We have also characterized the frequency of sister chromatid exchange (SCE) in transgenic Big Blue mouse cells. Big Blue Rat2 cells are hyperploid and have extremely high baseline frequencies of cytogenetic damage. In addition, Big Blue Rat2 cells are BrdU-resistant, therefore, SCE frequencies cannot be assessed in these cells. We conclude that Big Blue Rat2 cells are not useful for routine cytogenetic toxicology studies. The transgenic Big Blue mouse cell line is polyploid and consistently yields a low mitotic index (approximately 1%) in untreated cells. These mouse cells also exhibited moderately high baseline frequencies of chromosome and chromatid aberrations, however, baseline frequencies of SCE and of MN were not elevated. Transgenic Big Blue mouse embryonic fibroblasts were further studied for MN induction following treatment with N-ethyl-N-nitrosourea (ENU) for 0.5 h at concentrations of 0.425, 0.85 and 1.7 mM. Concentration-dependent increases in MN were observed in these cells. Thus, while an ENU-induced cytogenetic response using transgenic Big Blue mouse cells demonstrates that this cellular model could be used to cytogenetically complement the mutagenesis assays, the low mitotic index and the high spontaneous frequency of chromosome damage confounds its use for routine genetic toxicology studies.