Maintenance of bivalve hemocytes for the purpose of delayed DNA strand break assessment using the Comet assay

The lack of appropriate methods for storing intact and viable cells for the purpose of delayed DNA strand break analysis has hitherto limited the application of the Comet assay to in vitro or in vivo laboratory studies and restricted ecologically more relevant field-collected samples to sites in proximity to suitable laboratory facilities. In the present article, osmotically corrected cell culture media Hanks Balanced Salt Solution (HBSS) and Leibovitz Media (L-15) were assessed for their suitability as temporary storage media of blue mussel (Mytilus edulis) hemocytes. It was found that hemocytes maintained in either HBSS or L-15 could be stored for at least 7 days at 4°C without any significant deterioration in cell viability (Trypan blue) or increase in DNA strand breaks, expressed as % tail DNA. This approach allows the acquisition and examination of samples from organisms exposed in situ at previously unsuitable remote sites, thereby greatly increasing the potential ecological relevance of Comet assay-derived genotoxicity data. © 2009 Wiley-Liss, Inc.