细胞分离方法和药物浓度对固体肿瘤差异染色细胞毒性（DiSC）检测的影响。

Effect of cell isolation methods and drug concentration on the use of the Differential Staining Cytotoxicity (DiSC) assay with solid tumours.

机构信息

Bath Cancer Research Unit, Royal United Hospital, BA1 3NG, Bath, United Kingdom.

出版信息

Cytotechnology. 1989 Aug;2(3):225-32. doi: 10.1007/BF00133247.

Abstract

The Differential Staining Cytotoxicity (DiSC) in vitro drug sensitivity assay has been tested in solid tumours obtained from surgery. Various enzyme cocktails have been used for disaggregating cells for times ranging from 1-72 h. Collagenase (0.5%) plus DNase (0.002%) was chosen as producing the best results when incubated for 3 h at 37°C. This cocktail was also used for 24, 48 and 72 h incubations. Cell suspensions so produced were sometimes cleaned up by centrifugation on a Nycodenz gradient. Breast tumours were more often successfully cultured (67%) compared to gastrointestinal (43%) or other tumours (34%). Results with 6 drugs tested in 10 or more tumours suggested that in general, higher drug concentrations should be used when testing solid tumours as opposed to leukaemias in the DiSC assay.

摘要

体外药敏试验的差异染色细胞毒性（DiSC）已在手术获得的实体瘤中进行了测试。为了使细胞离散，使用了各种酶混合物，时间范围从 1 小时到 72 小时不等。当在 37°C 下孵育 3 小时时，选择胶原酶（0.5%）加 DNA 酶（0.002%）作为产生最佳结果的混合物。当孵育 24、48 和 72 小时时，也使用了这种混合物。如此产生的细胞悬浮液有时通过在 Nycodenz 梯度上离心进行清洗。与胃肠道（43%）或其他肿瘤（34%）相比，乳腺肿瘤的培养成功率更高（67%）。在 DiSC 测定中，用 6 种药物测试了 10 种或更多肿瘤的结果表明，与白血病相比，在测试实体瘤时，通常应使用更高的药物浓度。