INFLUENCE OF CULTURE PASSAGES ON GROWTH-KINETICS, XENOBIOTIC METABOLISM, CHROMOSOMAL STABILITY AND TRANSFORMATION IN A CLONAL FETAL HAMSTERLUNG EPITHELIAL-CELL LINE
C. Peiser et al., INFLUENCE OF CULTURE PASSAGES ON GROWTH-KINETICS, XENOBIOTIC METABOLISM, CHROMOSOMAL STABILITY AND TRANSFORMATION IN A CLONAL FETAL HAMSTERLUNG EPITHELIAL-CELL LINE, MUTATION RESEARCH, 289(2), 1993, pp. 281-290
M3E3/C3 is a clonal fetal hamster lung epithelial cell line which is u
sed for studies of epithelial differentiation as well as for in vitro
toxicologic tests. In this study growth kinetics, xenobiotic metabolis
m, chromosomal stability and transformation were investigated at incre
asing culture passage numbers up to 150. Cells of higher passages grew
faster and reached higher cell densities than the cells of lower ones
. As an indicator of xenobiotic metabolism we measured the activity of
7-ethoxycoumarin-deethylase (ECD), an enzyme belonging to the mixed f
unction oxidase system. Up to passage number 100 the ECD activity stro
ngly increased, followed by a slight decrease in additional passages.
The chromosomal stability was assessed by the induction of micronuclei
by benzo[a]pyrene (BaP) and N-methyl-N'-nitro-N-nitroso-guanidine (MN
NG). More micronuclei were always detected in cells of higher passages
than of lower passages. The capability of cells to be transformed to
anchorage independent growth by chemical carcinogens was examined usin
g a soft agar test. After carcinogen exposure with BaP and MNNG, cells
of higher passages showed higher transformation frequencies than cell
s of lower passages. Many cells at passage 150 exhibited an especially
high soft agar growth even without carcinogen treatment and were ther
efore characterized as spontaneously transformed. These results show t
hat metabolic and genetic characteristics of permanently growing cells
differ remarkably depending on the culture passage. This has always t
o be considered when permanently growing cells are used for toxicologi
cal studies.