Tetsuya T, Tetsuro K, Hiroshi Y, Touru T, Toshiharu F, Suguru K, Susumu K, Kyuya I, Osahiko A, Hirokazu Y
Dept. of Surgery, Keio University School of Medicine.
Gan To Kagaku Ryoho. 1991 Apr;18(4):547-54.
The chemosensitivity test with growth chamber (GC), a semi permeable polymer matrix, was conducted using human tumor xenografts, comparing the results with those of in vivo nude mouse system. Xenografts used were MX-1, St-4, Co-3, and Co-4. Normal stromal cells, SM-74, a cell line derived from human adult skin fibroblast, and Clone-A-31, a cell line from BALB/c nu/nu nude mice were used as the control. Dissociated tumor cell suspension in 200 microliters of Medium 199 was plated into GC (80,000 cells, chamber) and incubated with a various concentration of mitomycin C(MMC), cisplatin (DDP), 5-fluorouracil (5-FU), and adriamycin (ADM) at the various concentrations. After incubation of 1 wk, the activity of hexosaminidase was measured with ELISA assay using p-nitrophenyl-N-acetyl-glucosaminide. The antitumor activity of the agents against human tumor xenografts was dose dependent, and the antitumor spectra obtained by GC assay was essentially identical to in vivo nude mouse system. On the other hand, no evaluable optical density could be obtained with normal stromal cells, SM-74 and Clone-A-31. The optimal cutoff concentration of each drugs to predict the in vivo results was estimated to be 10 micrograms/ml for MMC, 15 micrograms/ml for DDP and 5-FU, and 0.7 microgram/ml for ADM. The predictability of GC assay was 77%, including 89% sensitivity and 70% specificity. Since GC assay could eliminate the normal stromal cells because of the characteristic of chamber surface, this assay was thought to be useful for the clinical chemosensitivity test of human cancers containing a large number of normal stromal cells.
使用生长室（GC）进行化学敏感性试验，生长室是一种半透性聚合物基质，采用人肿瘤异种移植模型，将结果与体内裸鼠系统的结果进行比较。所用的异种移植模型为MX-1、St-4、Co-3和Co-4。正常基质细胞SM-74（一种源自成人皮肤成纤维细胞的细胞系）和Clone-A-31（一种来自BALB/c nu/nu裸鼠的细胞系）用作对照。将200微升199培养基中的解离肿瘤细胞悬液接种到生长室（每个生长室80,000个细胞）中，并与不同浓度的丝裂霉素C（MMC）、顺铂（DDP）、5-氟尿嘧啶（5-FU）和阿霉素（ADM）在不同浓度下孵育。孵育1周后，使用对硝基苯基-N-乙酰氨基葡萄糖苷通过ELISA测定法测量己糖胺酶的活性。这些药物对人肿瘤异种移植模型的抗肿瘤活性呈剂量依赖性，通过生长室试验获得的抗肿瘤谱与体内裸鼠系统基本相同。另一方面，正常基质细胞SM-74和Clone-A-31未获得可评估的光密度。预测体内结果的每种药物的最佳截断浓度估计为MMC为10微克/毫升，DDP和5-FU为15微克/毫升，ADM为0.7微克/毫升。生长室试验的预测性为77%，包括89%的敏感性和70%的特异性。由于生长室试验因其腔室表面特性可消除正常基质细胞，因此该试验被认为对含有大量正常基质细胞的人类癌症的临床化学敏感性试验有用。
