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本文报道了用中国地鼠卵巢(CHO)细胞测试系统检测染色体畸变和姐妹染色单体交换(SCE)的方法。并经已知致癌物苯并(a)芘(BP)和黄曲霉毒素B_1验证。实验使用从国外引进的CHO细胞系,培养于最低基础培养液中,另补充非必需氨基酸和10％小牛血清,在含5％CO_2的培养箱中于37℃条件下培养。实验在加入和不加入肝匀浆S_9混合物的情况下,使受试物与细胞反应2小时。反应结束后24小时内收获细胞。作染色体分析用。如进行SCE分析用,则需于反应结束后加入含20μMBrdU的培养液。培养24～27小时,再加入秋水仙素。4小时后收获细胞。制片,用荧光素加姬姆萨染色法染色。 BP和黄曲霉毒素B_1诱发SCE频率和染色体畸变率的增加,并呈良好的剂量—反应关系。对肝匀浆S_9适宜浓度的研究表明,5％的浓度是合适的。
This article reports a method for detecting chromosomal aberrations and sister chromatid exchange (SCE) using the Chinese Hamster Ovary (CHO) Cell Assay System. And verified by the known carcinogens benzo (a) pyrene (BP) and aflatoxin B_1. Experiments using CHO cell lines introduced from abroad, cultured in minimal basal medium supplemented with nonessential amino acids and 10% fetal bovine serum in 5% CO 2 incubator at 37 ℃ under the conditions of culture. Experiments The subject was allowed to react with the cells for 2 hours with and without addition of a heparinized S_9 mixture. Cells were harvested within 24 hours after the end of the reaction. Used for chromosome analysis. For SCE analysis, add 20μM BrdU in the culture after the reaction. Cultivation of 24 to 27 hours, then add colchicine. Cells were harvested after 4 hours. Production, staining with fluorescein plus Giemsa staining. BP and aflatoxin B 1 induced an increase in SCE frequency and chromosomal aberration and showed a good dose-response relationship. Studies on the appropriate concentration of S_9 in liver homogenates have shown that a 5% concentration is appropriate.