PMID: 8591795Nov 1, 1995Paper

Characterizing mutagenesis in the hprt gene of rat alveolar epithelial cells

Experimental Lung Research
K E DriscollJ M Carter

Abstract

A clonal selection assay was developed for mutation in the hypoxanthine-guanine phosphoribosyl transferase (hprt) gene of rat alveolar epithelial cells. Studies were conducted to establish methods for isolation and long-term culture of rat alveolar epithelial cells. When isolated by pronase digestion purified on a Nycodenz gradient and cultured in media containing 7.5% fetal bovine serum (FBS), pituitary extract, EGF, insulin, and IGF-1, rat alveolar epithelial cells could be maintained in culture for several weeks with cell doubling times of 2-4 days. The rat alveolar epithelial cell cultures were exposed in vitro to the mutagens ethylnitrosourea (ENU) and H2O2, and mutation in the hprt gene was selected for by culture in the presence of the toxic purine analog, 6-thioguanine (6TG). In vitro exposure to ENU or H2O2 produced a dose-dependent increase in hprt mutation frequency in the alveolar epithelial cells. To determine if the assay system could be used to evaluate mutagenesis in alveolar type II cells after in vivo mutagen or carcinogen exposure, cells were isolated from rats treated previously with ENU or alpha-quartz. A significant increase in hprt mutation frequency was detected in alveolar epithelial cells obtained from...Continue Reading

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Citations

Aug 30, 2000·Toxicology in Vitro : an International Journal Published in Association with BIBRA·Z EliasB Fubini
May 30, 2002·Mutation Research·Roel P F SchinsPaul J A Borm
Dec 24, 1997·Environmental Health Perspectives·P J BormF J Schooten
Jul 16, 2005·Experimental Lung Research·Hugo Horlings, Peter Demant
Apr 12, 2001·Toxicology and Applied Pharmacology·L C RenwickA Clouter
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Oct 22, 2019·Journal of Toxicology and Environmental Health. Part B, Critical Reviews·Nicholas BirkettDaniel Krewski
Jul 1, 2004·Nonlinearity in Biology, Toxicology, Medicine·Bobby R ScottVernon E Walker

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