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一项涉及悬浮培养和软琼脂克隆的CHO/HGPRT突变试验评估:33种化学物质的结果

An evaluation of the CHO/HGPRT mutation assay involving suspension cultures and soft agar cloning: results for 33 chemicals.

作者信息

Oberly T J, Rexroat M A, Bewsey B J, Richardson K K, Michaelis K C

机构信息

Toxicology Division, Lilly Research Laboratories, Greenfield, Indiana 46140.

出版信息

Environ Mol Mutagen. 1990;16(4):260-71. doi: 10.1002/em.2850160408.

Abstract

The Chinese hamster ovary cell assay (CHO), which measures forward mutation of the HGPRT locus, is used in several laboratories for the detection of mutagens. A procedure involving treatment of CHO cells in suspension culture and mutant selection in soft agar cloning has been developed (Oberly TJ, Bewsey BJ, Probst GS (1987): Mutat Res 182:99-111). In order to evaluate the effectiveness of these modifications, 33 chemicals representing six chemical classes were tested, and the results were compared to findings obtained in other tests for genotoxicity at Lilly Research Laboratories (LRL). A positive response was obtained with 21 chemicals, all of which are recognized mutagens. Of the 12 compounds that produced negative results, 4 were considered to be mutagens and/or carcinogens. Twelve of the compounds mentioned in this report have been previously tested in the CHO/HGPRT assay by other laboratories, and the results showed strong agreement between laboratories. These findings support the conclusion that the use of suspension cultures and soft agar cloning in the CHO assay provides a sensitive test for the identification of mutagens and is a viable alternative to the traditional monolayer procedure of O'Neill et al. (O'Neill JP, Couch DB, Machanoff R, San Sebastian JR, Brimer PA, Hsie AW (1977): Mutat Res 45:103-109).

摘要

中国仓鼠卵巢细胞试验(CHO)用于检测诱变剂,该试验通过测量次黄嘌呤 - 鸟嘌呤磷酸核糖转移酶(HGPRT)位点的正向突变来进行。几个实验室采用了一种涉及悬浮培养中CHO细胞处理和软琼脂克隆中突变体选择的方法(奥伯利TJ、贝西BJ、普罗布斯特GS(1987年):《突变研究》182:99 - 111)。为了评估这些改进措施的有效性,对代表六类化学物质的33种化学物质进行了测试,并将结果与礼来研究实验室(LRL)其他遗传毒性测试的结果进行了比较。21种化学物质获得了阳性反应,所有这些物质均为公认的诱变剂。在产生阴性结果的12种化合物中,有4种被认为是诱变剂和/或致癌物。本报告中提到的12种化合物此前已由其他实验室在CHO/HGPRT试验中进行过测试,结果显示各实验室之间有很强的一致性。这些发现支持以下结论:在CHO试验中使用悬浮培养和软琼脂克隆为诱变剂的鉴定提供了一种灵敏的测试方法,并且是奥尼尔等人传统单层培养方法(奥尼尔JP、库奇DB、马查诺夫R、圣塞巴斯蒂安JR、布里默PA、谢AW(1977年):《突变研究》45:103 - 109)的可行替代方法。

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