SFTG international collaborative study on in vitro micronucleus test

Automation of the in vitro micronucleus assay using the Imagestream® imaging flow cytometer

“…Finally, mannitol (Fig. 5E), a substance that has been shown previously to be non‐cytotoxic and non‐genotoxic 14, induced a cytotoxicity of 30.1% at the highest dose of 4,000 μg/ml, but did not induce statistically significant increases in the frequency of MN compared with the solvent control at any doses.…”

“…Finally, for mannitol (Fig. 6E), the 3,000 μg/mL induced a cytotoxicity of 26.7% but no doses induced statistically significant increases in the frequency of MN when compared with solvent controls 14.…”

“…In recent years, several protocols and exposure schedules to perform the MN assay have been developed and validated in a number of recommended cell types 9, 10, 11, 12. Current forms of the assay are able to detect the presence of clastogens and aneugens in both rodent and human derived cell lines as well as in human peripheral blood lymphocytes 13, 14, 15, 16, 17.…”

Pesticide Biomarkers

“…When the damage occurs in the centromere, an imbalance in the distribution of the chromosomes is produced, disturbing the normal kinetics of anaphase and causing envelopment by a nuclear envelope. Assays of this type of damage may be performed using epithelial cells of the urinary bladder and oral and nasal mucosa (Stich & Rosin, 1984;Rosin & Gilbert, 1990) or peripheral blood (Lee et al, 2002;Clare et al, 2006). SCE occurs during the synthesis phase and represents symmetrical exchanges between homologous loci of replication products, occurring without DNA loss or changes in chromosome morphology (Norppa, 2004).…”

The use of an automated hematology analyzer to observe cell growth in the chromosome aberration test using human lymphocytes

“…One of the suggested reasons for this was that the mammalian cultured cell lines generally used lack the gene for p53 or the function of this gene is impaired, which causes the clastogenicity often associated with cytotoxicity without direct DNA damage (Hu et al, 1999;Galloway et al, 1998). However, assays using lymphocytes are considered to appropriately indicate the clastogenic potential of chemicals because they are derived from the living human body and possess the normal gene for p53 and repair system (Clare et al, 2006). However, it is difficult to observe or confirm the cell growth of lymphocytes, while it is straightforward to control that of cultured cell lines by seeding an exact number of cells at the start of culture and checking the growth by observing culture plates under a microscope or counting the cell number at the time of chemical treatment.…”