Kenyon N S, Schmittling R J, Siiman O, Burshteyn A, Bolton W E
Coulter Corporation, Miami, FL 33010.
Cytometry. 1994 Jun 1;16(2):175-83. doi: 10.1002/cyto.990160212.
Flow cytometry has become an important tool for the analysis of breast tumors, and assessment of S phase fraction and DNA ploidy are potential indicators of tumor aggression. Due to masking or dilution of infrequent tumor cell events, the presence of normal cell types, such as inflammatory cells and fibroblasts, can interfere with accurate DNA analysis of solid tumor samples. MDA-MB-175-VII human breast carcinoma cells, WI-38 human lung fibroblast cells, and peripheral blood leukocytes were mixed, in varying proportions, in order to represent human breast tumor samples. The cells were subsequently treated with CD45 conjugated magnetic microspheres to deplete tumor infiltrating leukocytes, thus enriching for tumor cells. The tumor cell mixtures were then stained with a pan-cytokeratin specific monoclonal antibody or with a monoclonal antibody that reacts with breast epithelial membrane antigen (EMA). When used in combination with monoclonal antibody gating, utilization of this bead-based technology resulted in enhanced precision of DNA analysis.
流式细胞术已成为分析乳腺肿瘤的重要工具，S期分数和DNA倍性评估是肿瘤侵袭性的潜在指标。由于罕见肿瘤细胞事件被掩盖或稀释，正常细胞类型（如炎性细胞和成纤维细胞）的存在会干扰实体瘤样本的准确DNA分析。将MDA-MB-175-VII人乳腺癌细胞、WI-38人肺成纤维细胞和外周血白细胞按不同比例混合，以代表人类乳腺肿瘤样本。随后用CD45偶联磁性微球处理细胞，以去除肿瘤浸润白细胞，从而富集肿瘤细胞。然后用全细胞角蛋白特异性单克隆抗体或与乳腺上皮膜抗原（EMA）反应的单克隆抗体对肿瘤细胞混合物进行染色。当与单克隆抗体门控结合使用时，这种基于磁珠的技术提高了DNA分析的精度。
