Campbell J A, Eppersimons C F, Kligerman A D, Petro A B, Sharief Y, Allen J W
In Vitro Cell Dev Biol. 1986 Aug;22(8):443-8. doi: 10.1007/BF02623444.
Methods are described for the short-term culture (48 to 56 h) of lung, liver, and kidney cells from C57B1/6 mice. With these techniques, mice can be exposed in vivo to test compounds and the cells grown on cover glasses in the presence of 5-bromo-2'-deoxyuridine (BrdUrd) (5 microM) for analysis of sister chromatid exchange (SCE) and cell cycle kinetics. Mice exposed to vinyl carbamate (VC) ((10 to 60 mg/kg) by i.p. injection were used in the initial examination of this system. Cultured lung and kidney cells from exposed animals (60 mg/kg) exhibited significant increases in SCE frequencies (approximately 3 to 5 times baseline); however, liver cells were much less responsive and showed less than a twofold increase over baseline SCE levels. Lung cultures initiated as long as 320 h after VC exposure (60 mg/kg) revealed a persistence of lesions leading to the formation of SCEs in vitro. This methodology permits analysis of cytogenetic damage in organs with very low mitotic activity after in vivo exposure to known or suspected genotoxicants.
本文描述了从C57B1/6小鼠获取肺、肝和肾细胞进行短期培养(48至56小时)的方法。利用这些技术,可使小鼠在体内接触受试化合物,然后将细胞培养在含有5-溴-2'-脱氧尿苷(BrdUrd)(5微摩尔)的盖玻片上,用于分析姐妹染色单体交换(SCE)和细胞周期动力学。通过腹腔注射给予小鼠乙烯基氨基甲酸酯(VC)(10至60毫克/千克),用于该系统的初步检测。暴露于VC(60毫克/千克)的动物所培养的肺和肾细胞,其SCE频率显著增加(约为基线的3至5倍);然而,肝细胞的反应则小得多,其SCE水平比基线增加不到两倍。在VC暴露(60毫克/千克)后长达320小时开始的肺培养显示,体外存在导致SCE形成的持续性损伤。该方法可用于分析体内接触已知或疑似遗传毒性物质后,有丝分裂活性极低的器官中的细胞遗传损伤。