Yang Y Y, Olofsson M, Köpf I, Hultborn R
Department of Oncology, Göteborg University, Sweden.
Anticancer Res. 1998 Jan-Feb;18(1A):53-9.
Clonogenic assays have been the golden standard for the assessment of cytotoxic injury from irradiation or drugs. Since such assays are time consuming, growth assays, often with automatic quantifying equipment, are frequently used. Since these procedures do not immediately reflect loss of clonogenic capacity, it was considered important to validate the two procedures using gamma-irradiation (0, 2 and 4 Gy) of two human cell lines (MCF-7 and Colo-205). The cells were growing exponentially in 96-well plates and crystal violet staining resulted in optical densities proportional to cell number. The homogeneity of optical densities within the plates was optimal if the wells to be measured were surrounded by liquid-containing ones. The slopes of the exponential growth curves were decreased upon irradiation. An "apparent cell survival", the mean of the three lowest ratios between irradiated and control cells, was defined. It was compared with the SF2 and SF4 as found in parallel Courtenay-Mills assays. In this work we found a modest underestimation of cell survival using the growth assay, ranging from 0 to 17 per cent in absolute terms.
克隆形成试验一直是评估辐射或药物引起的细胞毒性损伤的金标准。由于此类试验耗时较长,因此经常使用生长试验,通常配备自动定量设备。由于这些程序不能立即反映克隆形成能力的丧失,因此认为使用两种人类细胞系(MCF-7和Colo-205)进行γ射线照射(0、2和4 Gy)来验证这两种程序非常重要。细胞在96孔板中呈指数生长,结晶紫染色产生的光密度与细胞数量成正比。如果要测量的孔被含液体的孔包围,则板内光密度的均匀性最佳。照射后指数生长曲线的斜率降低。定义了“表观细胞存活率”,即照射细胞与对照细胞之间三个最低比率的平均值。将其与平行的考特尼-米尔斯试验中测得的SF2和SF4进行比较。在这项工作中,我们发现使用生长试验对细胞存活率的低估程度适中,绝对数值范围为0%至17%。