4 benzodiazepine sedatives, namely diazepam, medazepam, midazolam and bromazepam were investigated for their effects upon the fidelity of cell division in both low passage number and immortalised Chinese hamster cell lines. The study revealed substantial differences in the effect of these structurally related drugs upon mitosis, which may reflect different mechanisms of action of the drugs in cultured cells. Diazepam and medazepam exposure of immortal and low passage number cells resulted in the formation of monopolar mitotic spindles and subsequent metaphase arrest. The production of these spindles may be explained by the inhibition or centriole separation . In contrast, midazolam and bromazepam failed to produce observable changes in spindle structure. All 4 benzodiazepines produced significant toxicity in low passage number cells whereas, immortalised cells were more resistant to their toxic effects. They all induced metaphase chromosome dislocations in immortalised cells, whereas only diazepam and medazepam produced such effects in the low passage number cell line. In general, immortal cells appeared to be less sensitive to the toxic effects of benzodiazepines than the low passage number cells.