Ensley J F, Maciorowski Z, Pietraszkiewicz H, Hassan M, Kish J, Al-Sarraf M, Jacobs J, Weaver A, Atkinson D, Crissman J
Department of Internal Medicine, Wayne State University, Detroit, Michigan 48201.
Cytometry. 1987 Sep;8(5):488-93. doi: 10.1002/cyto.990080509.
Intense interest in advanced squamous cell cancers of the head and neck (SCC of H&N) has resulted from the recent progress made in tumor responses with chemotherapy and radiotherapy. Unfortunately, the response patterns and clinical outcome of such patients are not adequately predicted on an individual patient basis using clinical parameters or conventional morphology. The study of flow cytometrically determined cellular parameters in such tumors is therefore of interest, but is hindered by inadequate tumor preparative technology. The previous article (10) in this journal describes the use of a murine SCC tumor, LC12, which was employed for comparative testing and determination of optimum techniques of preparation for this tumor. This report describes the application of these techniques to 144 specimens of human SCC of H&N. The mean total yield for these specimens is 7.4 X 10(7) cells/g of tissue. The mean viable enzymatic yield (3.3 X 10(7) cells/g) was higher than the mean viable mechanical yield (2.0 X 10(7) cells/g) except when lymph nodes were the source of the specimen (5.4 X 10(7) cells/g). The mean dye exclusion viability from enzymatically dissociated specimens were above 90%. Significant aneuploidal subpopulation losses were evident in mechanically dissociated and enucleated specimens. 65% of the enzymatically dissociated specimens were successfully cultured with a mean cloning efficiency of 2.1 X 10(-3). Preparative techniques derived from comparative testing with a murine standard tumor have been successfully applied to 144 specimens of SCC of H&N with resultant high yields and excellent viability. Technical problems detected during the preliminary testing with LC12 were confirmed in the human tumors.
对头颈部晚期鳞状细胞癌（H&N SCC）的浓厚兴趣源于化疗和放疗在肿瘤反应方面最近取得的进展。不幸的是，使用临床参数或传统形态学方法，无法在个体患者基础上充分预测此类患者的反应模式和临床结果。因此，对流式细胞术测定此类肿瘤细胞参数的研究很有意义，但受到肿瘤制备技术不足的阻碍。本刊上一篇文章（10）描述了使用小鼠鳞状细胞癌肿瘤LC12，用于比较测试和确定该肿瘤的最佳制备技术。本报告描述了这些技术在144例人H&N SCC标本中的应用。这些标本的平均总细胞产量为7.4×10⁷个细胞/克组织。平均酶解活细胞产量（3.3×10⁷个细胞/克）高于平均机械法活细胞产量（2.0×10⁷个细胞/克），但当标本来源为淋巴结时除外（5.4×10⁷个细胞/克）。酶解标本的平均染料排除法活力高于90%。在机械解离和去核标本中明显存在显著的非整倍体亚群损失。65%的酶解标本成功培养，平均克隆效率为2.1×10⁻³。从与小鼠标准肿瘤的比较测试中得出的制备技术已成功应用于144例H&N SCC标本，从而获得了高产率和优异的活力。在对LC12的初步测试中检测到的技术问题在人类肿瘤中得到了证实。
