Premium
Improved sensitivity of the unscheduled DNA synthesis assay in primary rat hepatocytes following culture in serum‐free defined media
Author(s) -
Parton Joseph W.,
Yount David J.,
Garriott Michael L.
Publication year - 1995
Publication title -
environmental and molecular mutagenesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1
H-Index - 87
eISSN - 1098-2280
pISSN - 0893-6692
DOI - 10.1002/em.2850260208
Subject(s) - in vitro , in vivo , hepatocyte , in vitro toxicology , microbiology and biotechnology , biology , dna , chemistry , biochemistry , pharmacology
The unscheduled DNA synthesis (UDS) assay has been used extensively for the in vitro detection of DNA damage caused by compound exposure. However, the in vitro UDS assay has been insensitive for the detection of certain chemicals, particularly nitroaromatic compounds, that are positive in bacterial mutation assays. Recently, studies have been reported which describe alterations in the he‐patocyte membrane following collagenase perfusion. Independently, a method for serum‐free tissue culture has been developed which results in the up‐regulation of cell surface receptors and which may restore membrane functions. Fourteen compounds, including seven nitroaromatics, were evaluated in the in vitro UDS assay employing a serum‐free procedure. Five compounds that were previously reported positive in the standard in vitro UDS assay were also found positive using the serum‐free method. In addition, five of the nitroaromatic compounds produced positive results with the serum‐free method. 1‐Methyl‐3‐nitro‐1‐nitrosoguanidine and 2‐acetylaminofluorene, routinely used as positive controls in the UDS assay, showed greater activity in the serum‐free assay. These results suggest that the use of serum‐free media improves the sensitivity of the in vitro UDS assay and that the serum‐free procedure potentially offers an effective alternative to the more labor intensive and more costly in vivo UDS assay. © 1995 Wiley‐Liss, Inc.