Friedman H M, Glaubiger D L
Cancer Res. 1982 Nov;42(11):4683-9.
We have developed a method for performing in vitro drug testing on primary human tumor explants which is a variant of the human tumor stem cell assay (HTSCA) described previously by Salmon et al. (N. Engl. J. Med. 298: 1321, 1978). The method utilizes a cell-containing liquid top layer and a soft-agar bottom layer. Tumor growth is measured by [3H]thymidine incorporation into material precipitable by 5% trichloroacetate. Results show linear correlations with number of cells plated and with a number of colonies per plate measured using the HTSCA, when cell aliquots from one sample are used. In vitro drug sensitivity, as determined by inhibition of [3H]thymidine incorporation, correlates with HTSCA results in 54 of 61 determinations (89%). Of 22 experiments in which drug sensitivity curves were compared, 21 (95%) were similar in both systems. The [3H]thymidine method yields results more quickly (5 days after samples are plated) and with smaller variances than those measurements obtained using the HTSCA. Normal human skin muscle, lung, and colon tissue and a human fibroblast cell line do not incorporate significant amounts of [3H]thymidine into trichloroacetic acid-precipitable material. Thus, normal cell components plated in tumor samples do not interfere with assay results. Standard scintillation counting is used; optical counting, either visual or automated, is not required. Therefore, the measurement of in vitro drug sensitivity by inhibition of incorporation of radiolabeled precursors deserves further evaluation as a predictor of in vitro response.
我们开发了一种在原发性人肿瘤外植体上进行体外药物测试的方法，该方法是Salmon等人（《新英格兰医学杂志》298: 1321, 1978）之前描述的人肿瘤干细胞测定法（HTSCA）的一种变体。该方法利用含细胞的液体顶层和软琼脂底层。通过将[³H]胸腺嘧啶核苷掺入可被5%三氯乙酸沉淀的物质中来测量肿瘤生长。当使用来自一个样本的细胞等分试样时，结果显示与接种的细胞数量以及使用HTSCA测量的每平板菌落数量呈线性相关。通过抑制[³H]胸腺嘧啶核苷掺入所确定的体外药物敏感性，在61次测定中有54次（89%）与HTSCA结果相关。在22次比较药物敏感性曲线的实验中，两个系统中有21次（95%）相似。[³H]胸腺嘧啶核苷方法产生结果的速度更快（接种样本后5天），并且方差比使用HTSCA获得的测量结果更小。正常人皮肤、肌肉、肺和结肠组织以及人成纤维细胞系不会将大量的[³H]胸腺嘧啶核苷掺入三氯乙酸可沉淀物质中。因此，接种在肿瘤样本中的正常细胞成分不会干扰测定结果。使用标准闪烁计数；不需要视觉或自动的光学计数。因此，通过抑制放射性标记前体掺入来测量体外药物敏感性作为体外反应的预测指标值得进一步评估。
