A method for in vitro culture of rat Zymbal gland: use in mechanistic studies of benzene carcinogenesis in combination with 32P-postlabeling

Environmental Health Perspectives
M V ReddyM A Mehlman

Abstract

Zymbal glands were excised bilaterally from the ear ducts of female Sprague-Dawley rats (three/group), minced into approximately four fragments per gland, and transferred into a microtiter plate containing 1.5 mL per well of Waymouth's tissue culture medium supplemented with fetal calf serum, hydrocortisone, insulin, and gentamicin. After addition of a test compound or solvent vehicle, plates were incubated for 6, 24, 48, or 96 hr at 37 degrees C in a humidified atmosphere of 5% CO2 in air. Tissue in culture for 6 hr was histologically indistinguishable from the freshly excised tissue, while that in culture for 24, 48, and 96 hr showed a progressive deterioration often with necrosis and/or squamous metaplasia. More pronounced deterioration was noted in samples treated with 750 or 1500 micrograms/mL of benzene. Using a nuclease P1-enhanced 32P-postlabeling assay, aromatic DNA adducts were detected in cultured Zymbal glands exposed for 48 hr to benzene and its derivatives, as well as to 7,12-dimethylbenzanthracene (DMBA) and 2-acetylaminofluorene (AAF). Benzene produced very low levels of adducts (0.5 adducts per 10(9) nucleotides), whereas its congeners produced relatively high levels of adducts (50-2000 lesions per 10(9) nucleo...Continue Reading

References

Jul 1, 1989·Environmental Health Perspectives·M V ReddyC R Mackerer
Jan 1, 1987·Critical Reviews in Toxicology·G F Kalf
Jul 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·J G LiehrK Randerath
Nov 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·L LatrianoG Witz
Oct 1, 1985·Environmental Health Perspectives·K RanderathM V Reddy
Nov 1, 1981·Analytical Biochemistry·M V ReddyK Randerath
Jan 1, 1981·Journal of Supramolecular Structure and Cellular Biochemistry·I B Weinstein

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Citations

Dec 1, 1994·Mutation Research·M V ReddyC R Mackerer
Jul 1, 1989·Environmental Health Perspectives·M V ReddyC R Mackerer
Apr 1, 1993·Environmental Health Perspectives·R SnyderB D Goldstein
Dec 1, 1996·Environmental Health Perspectives·K H NorpothE Jorg
Jun 24, 1999·Critical Reviews in Toxicology·A P DeCaprio
Dec 1, 1996·Environmental Health Perspectives·F A AngelosantoC R Mackerer

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