Differential sensitivity of Chinese hamster V79 and Chinese hamster ovary (CHO) cells in the in vitro micronucleus screening assay.
Both the V79 and CHO cell lines are routinely used in the in vitro MN screening assay for the detection of possible genotoxicants. The CHO cell line is the predominant cell line currently used in the genetic toxicology testing industry. However, some laboratories routinely utilize the V79 cell line since the in vitro MN screening assay was initially developed using V79 cells. Our laboratory has historically used the CHO cell line. Therefore, our laboratory was interested in comparing the two cell lines with regard to possible similarities or differences in MN induction sensitivity after exposure to cyclophosphamide (CPA) and mitomycin C (MMC), the two standard positive control chemicals routinely used in this assay. Three exposure conditions in the presence of CPA and MMC were examined in both cell lines. Replicate cultures of CHO cells in McCoy's 5A and V79 cells in both McCoy's 5A and E-MEM were established and treated with 5 microg CPA/ml (4h exposure with S9), 0.5 microg MMC (4h exposure without S9) and 0.5 microg MMC (24h exposure without S9). A total of 400 cytochalasin B-blocked binucleated cells and 200 consecutive cells were analyzed from each culture for MN and cell cycle kinetics, respectively. Analysis of the data demonstrated that CHO cells were up to approximately five-fold more sensitive to the induction of CPA- and MMC-induced MN than V79 cells. Both cell lines exhibited similar average generation times among identical exposure groups. Therefore, the difference in MN sensitivity cannot be attributed to possible differences in cell cycle kinetics and is possibly related to inherent cellular differences in the processing of and/or repair of CPA- and MMC-induced damage by V79 and CHO cells.
Erexson
G L
GL
Covance Laboratories, Inc., Department of Genetic and Molecular Toxicology, 9200 Leesburg Pike, Vienna, VA 22182, USA. gregory.erexson@covance.com
Periago
M V
MV
Spicer
C S
CS
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D003520:Cyclophosphamide / Q000633:toxicity*; D015162:Micronucleus Tests; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology; D016685:Mitomycin / Q000633:toxicity*; D009153:Mutagens / Q000633:toxicity*; D012680:Sensitivity and Specificity
D009153:Mutagens; D016685:Mitomycin; D003520:Cyclophosphamide