Premium
A rapid in vitro assay for carcinogenicity of chemical substances in mammalian cells utilizing an attachment‐independence endpoint
Author(s) -
Traul K. A.,
Kachevsky V.,
Wolff J. S.,
Hink R.,
Nanna U.
Publication year - 1979
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.2910230209
Subject(s) - carcinogen , in vitro , petri dish , chemistry , pyrene , agar , cell culture , cell type , biology , cell , microbiology and biotechnology , biochemistry , genetics , organic chemistry , bacteria
Rauscher leukemia virus (RLV)‐infected rat embryo cells have been shown to be sensitive to neoplastic transformation by a variety of chemical carcinogens. The mass assay technique normally used requires 6 to 15 weeks before morphologically altered foci of cells are observed. While growth in soft agar is considered to be one of the best in vitro means for confirming neoplastic transformation, this assay requires 4 additional weeks beyond focus formation. We have devised a rapid new assay for determining carcinogenicity of chemicals. The method takes advantage of the range of chemical sensitivity of RLV‐infected rat embryo cells, but provides, in 10 days, for a readout based on the ability of transformed cells to survive under conditions which select for attachment independence. The target cells, 2FR 4 50, were treated for 72 h with chemical and then plated into Petri dishes containing a solid bottom layer of agar/medium. Viable cell counts were made 3 and 6 days later and compared to parallel data from solvent‐treated control cells. 2FR 4 50 cells treated with any of 10 chemical carcinogens, including such diverse compounds as benzo( a )pyrene, 3‐methylcholanthrene, 2‐acetylaminofluorene, NiSO 4 and urethane, showed an average increase of 225% in survival over controls. Phenanthrene, a non‐carcinogen, induced no increase in survival of 2FR 4 50 cells. This assay appears to permit detection of carcinogens of various chemical types with a simple test readout within 10 days.