Slocum H K, Pavelic Z P, Rustum Y M, Creaven P J, Karakousis C, Takita H, Greco W R
Cancer Res. 1981 Apr;41(4):1428-34.
Characterization of cells comprising solid tumors will facilitate the rational design of cancer chemotherapy for individual patients. We have prepared cell suspensions from human melanoma, sarcoma, and lung tumors by thinly slicing the tissue with a microtome and scalpels (mechanical release), followed by treatment with a mixture of collagenase II and DNase I (enzymatic release). This method of disaggregation resulted in two cell suspensions for each tumor specimen, and we characterized these suspensions by assessing their dye exclusion capability, ribonucleoside triphosphate pools, cytological profile and clonogenicity in soft agar. The enzymatic method thus yields cells in addition to those obtainable by a mild mechanical procedure, and these cells are similar in cytological profile and clonogenicity in soft agar to those released mechanically. Furthermore, the enzymatically released population is superior to that released mechanically for purposes requiring large numbers of dye-excluding cells having intact ribonucleotide pools.
对构成实体瘤的细胞进行表征将有助于为个体患者合理设计癌症化疗方案。我们通过用切片机和手术刀将组织切成薄片(机械释放),然后用胶原酶II和脱氧核糖核酸酶I的混合物处理(酶促释放),从人黑色素瘤、肉瘤和肺癌肿瘤中制备了细胞悬液。这种解离方法为每个肿瘤标本产生了两种细胞悬液,我们通过评估它们的染料排斥能力、三磷酸核糖核苷池、细胞学特征和在软琼脂中的克隆形成能力来表征这些悬液。因此,酶促方法除了能产生通过温和机械程序获得的细胞外,还能产生其他细胞,这些细胞在细胞学特征和在软琼脂中的克隆形成能力方面与机械释放的细胞相似。此外,对于需要大量具有完整核糖核苷酸池的染料排斥细胞的目的,酶促释放的细胞群体优于机械释放的细胞群体。