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A method for detecting carcinogenic organic chemicals using mammalian cells in culture.

机译:一种使用培养的哺乳动物细胞检测致癌有机化学物质的方法。

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摘要

A method for testing organic chemicals for their carcinogenic potential is described. Baby hamster kidney cells (BHK-21/C1 13) were exposed to different doses of test compound in liquid tissue culture medium containing rat liver post-mitochondrial supernatant and cofactors (S-9 mix) to aid metabolism, but without serum. Survival of cells following exposure to the compound was assessed by cloning in liquid growth medium. Transformation was assessed by colony growth in semi-solid agar. The dose-response curve for survival was used to determine the LC50 of the compound. A dose-response curve for transformation was constructed and a 5-fold increase in transformation frequency at the LC50 was regarded as a positive test result. The method may also be used for testing gaseous compounds. Cells grown in monolayers and overlaid with serum-free medium and S-9 mix were exposed to vinyl chloride gas mixed with air. After exposure, the treated cells were trypsinized, resuspended in growth medium, and survival and transformation assays performed. The methods described are illustrated by examples taken from an evaluation study using 120 compounds and found to be more than 90% accurate in distinguishing between carcinogens and non-carcinogens.
机译:描述了一种测试有机化学物质潜在致癌性的方法。在含有大鼠肝线粒体后上清液和辅助因子(S-9混合物)的液体组织培养基中,将幼仓鼠肾细胞(BHK-21 / C1 13)暴露于不同剂量的测试化合物,以帮助代谢,但不需血清。通过克隆在液体生长培养基中来评估暴露于化合物后的细胞存活率。通过半固体琼脂中菌落生长评估转化。存活的剂量反应曲线用于确定化合物的LC50。构建了转化的剂量反应曲线,LC50转化频率的5倍增加被认为是阳性测试结果。该方法也可以用于测试气态化合物。将单层生长并覆盖无血清培养基和S-9混合物的细胞暴露于与空气混合的氯乙烯气体中。暴露后,将经处理的细胞用胰蛋白酶消化,重悬于生长培养基中,并进行存活和转化测定。所描述的方法通过使用120种化合物进行的评估研究中的实例进行了说明,发现在区分致癌物和非致癌物方面准确度超过90%。

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