Dayeh Vivian R, Chow Stephanie L, Schirmer Kristin, Lynn Denis H, Bols Niels C
Department of Biology, University of Waterloo, Waterloo, Ont, Canada N2L 3G1.
Ecotoxicol Environ Saf. 2004 Mar;57(3):375-82. doi: 10.1016/S0147-6513(03)00083-6.
Three viability assays using fluorescent dyes effectively detected a loss of viability in cultures of three mammalian cell lines (H4IIE, Caco2, and HepG-2), two fish cell lines (RTgill-W1 and RTL-W1), and a ciliated protozoan, Tetrahymena thermophila, after exposure to Triton X-100, used as a model toxicant. The dyes were Alamar Blue (AB), neutral red (NR), and propidium iodide, which respectively monitored energy metabolism, lysosomal activity, and membrane integrity. A fourth fluorescent dye, 5-carboxyfluorescein diacetate acetoxymethyl ester, was problematic. For 2-h Triton X-100 exposures, mammalian cell lines were as susceptible as piscine cell lines, whereas T. thermophila was approximately twofold less sensitive as detected with AB and NR. Despite being less sensitive, cytotoxicity tests on T. thermophila could be done in spring water, which means that unlike animal cells they could be directly exposed to most industrial effluents without osmolality adjustments. Therefore, T. thermophila could be a useful complement to animal cells as alternatives to fish in toxicity testing.
使用荧光染料的三种活力测定法有效地检测到，在暴露于用作模型毒物的 Triton X-100 后，三种哺乳动物细胞系（H4IIE、Caco2 和 HepG-2）、两种鱼类细胞系（RTgill-W1 和 RTL-W1）以及一种纤毛原生动物嗜热四膜虫的培养物中活力丧失。这些染料分别是阿拉玛蓝（AB）、中性红（NR）和碘化丙啶，它们分别监测能量代谢、溶酶体活性和膜完整性。第四种荧光染料 5-羧基荧光素二乙酸酯乙酰氧基甲酯存在问题。对于 2 小时的 Triton X-100 暴露，哺乳动物细胞系与鱼类细胞系一样敏感，而用 AB 和 NR 检测时，嗜热四膜虫的敏感性约低两倍。尽管敏感性较低，但对嗜热四膜虫的细胞毒性测试可以在泉水中进行，这意味着与动物细胞不同，它们可以直接暴露于大多数工业废水中而无需进行渗透压调节。因此，嗜热四膜虫作为毒性测试中鱼类的替代品，可能是动物细胞的有用补充。
