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首页> 外文期刊>Cytometry, Part A: the journal of the International Society for Analytical Cytology >Detection and quantification of circulating tumor cells in mouse models of human breast cancer using immunomagnetic enrichment and multiparameter flow cytometry
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Detection and quantification of circulating tumor cells in mouse models of human breast cancer using immunomagnetic enrichment and multiparameter flow cytometry

机译：使用免疫磁富集和多参数流式细胞术检测和定量人类乳腺癌小鼠模型中循环肿瘤细胞

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Background: Circulating tumor cells (CTCs) in the peripheral blood of breast cancer patients may be an important indicator of metastatic disease and poor prognosis. However, the use of experimental models is required to fully elucidate the functional consequences of CTCs. The purpose of this study was to optimize the sensitivity of multiparameter flow cytometry for detection of human tumor cells in mouse models of breast cancer.Methods: MDA-MB-468 human breast cancer cells were serially diluted in whole mouse blood. Samples were lysed and incubated with a fluorescein isothiocyanate-conjugated anti-human leukocytic antigen antibody and a phycoerythrin-conjugated anti-mouse pan-leukocyte CD45 antibody. Samples were then immunomagnetically depleted of CD45-positive leukocytes, fixed, permeabilized, and stained with propidium iodide before flow cytometric analysis.Results: Human breast cancer cells could be differentiated from mouse leukocytes based on increased light scatter, cell surface marker expression, and aneuploid DNA content. The method was found to have a lower sensitivity limit of 10(-5) and was effective for detecting human breast cancer cells in vivo in the circulation of experimental mice carrying primary human mammary tumors.Conclusions: This technique has the potential to be a valuable and sensitive tool for investigating the biological relevance of CTCs in experimental mouse models of breast cancer. (c) 2005 Wiley-Liss, Inc.
机译：背景：乳腺癌患者外周血中的循环肿瘤细胞（CTC）可能是转移性疾病和预后不良的重要指标。但是，需要使用实验模型来充分阐明CTC的功能后果。这项研究的目的是优化多参数流式细胞仪检测乳腺癌小鼠模型中人肿瘤细胞的敏感性。方法：在小鼠全血中连续稀释MDA-MB-468人乳腺癌细胞。裂解样品，并与结合异硫氰酸荧光素的抗人白细胞抗原抗体和结合藻红蛋白的抗小鼠泛白细胞CD45抗体一起孵育。然后在流式细胞仪分析之前，对样本中的CD45阳性白细胞进行免疫磁化处理，固定，通透并用碘化丙啶染色。结果：根据光散射，细胞表面标志物表达和非整倍体的增加，可以将人乳腺癌细胞与小鼠白细胞区分开来。 DNA含量。该方法的灵敏度下限为10（-5），可有效检测体内携带人原发性乳腺肿瘤的实验小鼠体内的人乳腺癌细胞。结论：该技术具有潜在的实用价值和敏感性工具，用于研究乳腺癌实验小鼠模型中四氯化碳的生物学相关性。 （c）2005 Wiley-Liss，Inc.

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