Oberly T J, Rexroat M A, Richardson K K
Toxicology Division, Lilly Research Laboratories, Eli Lilly and Company, Greenfield, IN 46140.
Mutat Res. 1990 Jun;244(2):105-9. doi: 10.1016/0165-7992(90)90057-q.
Iron-supplemented bovine calf serum (ICS) was found to be a viable alternative to fetal bovine serum (FBS) in the growth promotion and cloning efficiency of Chinese hamster ovary (CHO) cells that are used in the HGPRT mutation assay. Suspension cultures of CHO cells had an average generation time of 11.5 h in ICS and 13.6 h for cells maintained in FBS. This slight difference was due to lot variability on the part of FBS and could be eliminated by routine quality control measures. The average cloning efficiencies for CHO cells cloned in either ICS or FBS were 107% and 88%, respectively, and these values were not statistically different. No appreciable difference was noted in the spontaneous mutation rates of cells cloned in either ICS or FBS. Furthermore, the use of ICS in mutagenicity studies with genotoxic agents shows the serum to be at least equal or superior to FBS in the detection of both direct-acting mutagens and promutagens. These data suggest that ICS is an appropriate serum to be used in the CHO/HGPRT test system. Since ICS is more readily available and considerably less costly than FBS, a substantial reduction in the cost of the assay can be realized.
在用于次黄嘌呤鸟嘌呤磷酸核糖转移酶(HGPRT)突变试验的中国仓鼠卵巢(CHO)细胞的生长促进和克隆效率方面,发现补充铁的小牛血清(ICS)是胎牛血清(FBS)的可行替代品。CHO细胞的悬浮培养物在ICS中的平均代时为11.5小时,而在FBS中培养的细胞平均代时为13.6小时。这种细微差异是由于FBS批次的变异性造成的,可通过常规质量控制措施消除。在ICS或FBS中克隆的CHO细胞的平均克隆效率分别为107%和88%,这些值在统计学上没有差异。在ICS或FBS中克隆的细胞的自发突变率没有明显差异。此外,在遗传毒性剂的致突变性研究中使用ICS表明,在检测直接作用诱变剂和前诱变剂方面,该血清至少与FBS相当或优于FBS。这些数据表明ICS是CHO/HGPRT测试系统中使用的合适血清。由于ICS比FBS更容易获得且成本低得多,因此可以大幅降低试验成本。