O'Neill J P, Sullivan L M, Booker J K, Pornelos B S, Falta M T, Greene C J, Albertini R J
Vermont Regional Cancer Center, University of Vermont, Burlington 05401.
Environ Mol Mutagen. 1989;13(4):289-93. doi: 10.1002/em.2850130403.
The in vivo frequency of mutants resulting from mutation at the hprt locus in human T-lymphocytes can be determined by a cloning assay. This assay quantifies the frequency of 6-thioguanine-resistant (TGr) T-cells through growth of colonies in 96-well microtiter dishes. The reproducibility of the TGr mutant frequency values has now been assessed in a longitudinal study of six individuals (three male, three female, aged 22-33 years) employing 4-5 blood samples over a 26-37 week time period. Cloning assays were performed with both fresh and cryopreserved cell samples. No significant differences were found among the mutant frequency values for multiple samples from each individual with both fresh and cryopreserved cell samples. These results demonstrate the reproducibility of this cloning assay for in vivo mutant frequency determinations in human T-lymphocytes.
通过克隆试验可以确定人类T淋巴细胞中次黄嘌呤-鸟嘌呤磷酸核糖转移酶（hprt）基因座突变产生的突变体的体内频率。该试验通过在96孔微量滴定板中培养菌落来量化6-硫鸟嘌呤抗性（TGr）T细胞的频率。现在，在一项纵向研究中评估了6名个体（3名男性，3名女性，年龄在22-33岁之间）的TGr突变频率值的可重复性，该研究在26-37周的时间段内采集了4-5份血样。使用新鲜和冷冻保存的细胞样本进行克隆试验。对于来自每个个体的多个新鲜和冷冻保存细胞样本的突变频率值，未发现显著差异。这些结果证明了这种克隆试验在测定人类T淋巴细胞体内突变频率方面的可重复性。
