Permanent embryonic germ cell lines of BALB/cJ mice--an in vitro alternative for in vivo germ cell mutagenicity tests.
To offer a sensitive and predictive in vitro method to assess germ cell mutagenicity, we established primordial germ (PG) cell-derived permanent female and male embryonic germ (EG) cell lines of the mouse (strain BALB/cJ). The differences in developmental sensitivity of EG cells and differentiated fibroblast cells of the mouse cell line 3T3 to genotoxicants were tested comparatively under identical test conditions. Cytotoxicity assay was measured by the MTT test and genotoxic effects were determined by sister chromatid exchanges (SCE) rates induced by standard reference mutagens. Both methods are used to assign the chemicals to two classes of in vivo reproductive toxicity, non- and strongly genotoxic to germ cells. Applying linear discriminant analysis, a biostatistical prediction model (PM) was developed for the female cell line EG(3). This procedure identified a single variable, the Ig(SCE(200)EG(3)) as the statistically significant concentration related increase of 200% in the mean number of SCEs per metaphase spread after 3 h of exposure to be sufficient for separation into the classes: non- and strongly genotoxic to germ cells. Applying this PM to the training set of five genotoxic and three non-genotoxic test chemicals, 100% correct classifications were obtained.
Klemm
M
M
Center for Documentation and Evaluation of Alternative Methods to Animal Experiments (ZEBET), Federal Institute for Health Protection of Consumers and Veterinary Medicine (BgVV), Berlin, Germany.
Genschow
E
E
Pohl
I
I
Barrabas
C
C
Liebsch
M
M
Spielmann
H
H
D016475:3T3 Cells; D000826:Animal Testing Alternatives; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D016002:Discriminant Analysis; D004305:Dose-Response Relationship, Drug; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D005854:Germ Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D015233:Models, Statistical; D009152:Mutagenicity Tests / Q000379:methods*; D009153:Mutagens / Q000633:toxicity*; D011237:Predictive Value of Tests; D012732:Sex Determination Analysis; D012854:Sister Chromatid Exchange / Q000187:drug effects; D013778:Tetrazolium Salts / Q000378:metabolism
D009153:Mutagens; D013778:Tetrazolium Salts