Effects of Different Polycyclic Aromatic Hydrocarbons on Cultured Fetal Hamster Lung Cells and Tracheal Explants
A number of polycyclic aromatic hydrocarbons have been suspected of playing a role in human pulmonary carcinogenesis. In order to test the biologic effects of pure single compounds we have developed a new test system which combines a cell transformation and an organ culture assay.
Short-term cultured lung cells, isolated from fetuses which were dissected aseptically from pregnant female Syrian golden hamsters at the 15th day of gestation, were plated at a density of 3–6×103 into 60 mm Petri dishes. Test substances dissolved in not more than 0.5% DMSO were added to the cells 1 day after plating; 24 h later, the cultures were washed three times with Hanks’ solution and re-fed with normal medium. After 10–14 days, the cultures were fixed with methanol and Giemsa stained.
The evaluation showed a dose-related toxicity of all tested compounds (i.e., benzo(a)pyrene, benz(a)anthracene, benzo(e)pyrene, and chrysene). Transformation could be obviously detected with benzo(a)pyrene, while the other test compounds induced only weak or no transformation.
Tracheal pieces from the same fetuses were dissected and transferred to organ culture. Morphological alterations caused by the same test compounds were examined on serial sections of the explants after a cultivation time of 4–8 weeks. The exposure to chemicals started 1 day after the beginning of cultivation and lasted 1 week.
As this system has been initiated very recently, the results are only preliminary and have to be improved upon. Our primary concern has been directed at the consistency of correlations between the results of lung cell and tracheal explant cultures. Therefore, the feasibilities for standardizing the in vitro conditions as well as the advantages of adopting the system for other tissues (human) will be discussed.
KeywordsCellulose Toxicity Agar DMSO Hydrocarbon
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