Taetle R, Honeysett J M, Rosen F, Shoemaker R
Cancer. 1986 Nov 1;58(9):1969-78. doi: 10.1002/1097-0142(19861101)58:9<1969::aid-cncr2820580903>3.0.co;2-4.
Previous studies suggested tumor colony-forming cells (CFC) grown from xenografts might be useful as a preclinical, in vitro drug screen. To further evaluate this possibility, eight melanoma and six ovarian carcinoma xenografts were established from untreated patients and tested for in vitro CFC growth. For each tumor, linear relationships between cells plated and colony (30 cells or greater than 75 micron diameter) and cluster (10-30 cells or 50-75 micron) growth were observed. All eight melanomas grew sufficient colonies (greater than or equal to 30) for in vitro drug assessment, although four required hypoxic (pO2 = 40) incubation to reliably attain this level of growth. Only one in six of the ovary xenografts consistently grew enough colonies, and growth was not significantly improved by hypoxic incubation, or addition of luteinizing hormone, follicle-stimulating hormone, or steroid hormones. Cloning efficiencies (colonies + clusters/cells plated) for tumors demonstrating adequate growth ranged from 0.01% to 0.3%. For most tumors, no direct relationship was observed between characteristics of xenograft tumors (size) or their resulting cell suspensions (viabilities, cell yield) and CFC growth. Cell suspensions were incubated with a 3 log concentration of nine established chemotherapeutic agents. Resulting dose-effect curves were linear and showed no plateaus of drug effect. Analyzing 447 in vitro drug trials on six melanomas and one ovarian carcinoma, interexperiment variability was high. Cell lines were established from three xenografts using a low concentration of fetal bovine serum (1%), and also examined for in vitro drug sensitivity. Using both liquid culture isotope incorporation and a colony-forming assay, drug sensitivity profiles for the cell lines were nearly identical to those for parent xenograft CFC. However, assays performed using the cell lines were more reproducible than those using xenograft tissue. The authors conclude that tumor CFC can be reliably grown from melanoma xenografts, but in vitro drug assays using these xenografts are poorly reproducible. The xenografts are a resource for establishing cell lines, and drug assays performed using these lines are highly reproducible. Similarities in drug sensitivity profiles for parent xenograft CFC and derived cell lines suggest that, despite poor reproducibility, repetitive assays using melanoma CFC accurately reflect some properties of cells which sustain tumor cell growth.
先前的研究表明，从异种移植瘤中培养出的肿瘤集落形成细胞（CFC）可能有助于进行临床前的体外药物筛选。为了进一步评估这种可能性，从未经治疗的患者中建立了8个黑色素瘤和6个卵巢癌异种移植瘤，并对其进行体外CFC生长测试。对于每个肿瘤，观察到接种细胞与集落（30个细胞或直径大于75微米）和细胞簇（10 - 30个细胞或50 - 75微米）生长之间呈线性关系。所有8个黑色素瘤都生长出了足够数量（大于或等于30个）的集落用于体外药物评估，尽管其中4个需要在低氧（pO2 = 40）条件下培养才能可靠地达到这种生长水平。6个卵巢癌异种移植瘤中只有1个始终能生长出足够数量的集落，低氧培养、添加促黄体生成素、促卵泡生成素或类固醇激素均未显著改善其生长情况。生长良好的肿瘤的克隆效率（集落 + 细胞簇/接种细胞数）在0.01%至0.3%之间。对于大多数肿瘤，未观察到异种移植瘤的特征（大小）或其产生的细胞悬液（活力、细胞产量）与CFC生长之间存在直接关系。将细胞悬液与9种已确定化疗药物的3个对数浓度进行孵育。所得剂量 - 效应曲线呈线性，且未显示出药物效应的平台期。分析对6个黑色素瘤和1个卵巢癌进行的447次体外药物试验，实验间变异性较高。使用低浓度胎牛血清（1%）从3个异种移植瘤中建立细胞系，并对其进行体外药物敏感性检测。使用液体培养同位素掺入法和集落形成试验，细胞系的药物敏感性谱与亲本异种移植瘤CFC的几乎相同。然而，使用细胞系进行的检测比使用异种移植组织进行的检测更具可重复性。作者得出结论，肿瘤CFC可以从黑色素瘤异种移植瘤中可靠地培养出来，但使用这些异种移植瘤进行的体外药物检测重复性较差。异种移植瘤是建立细胞系的资源，使用这些细胞系进行的药物检测具有高度可重复性。亲本异种移植瘤CFC和衍生细胞系的药物敏感性谱相似，这表明尽管重复性较差，但使用黑色素瘤CFC进行的重复检测准确反映了维持肿瘤细胞生长的细胞的某些特性。
