Flow cytometric enrichment for respiratory epithelial cells in sputum
Flow cytometric enrichment for respiratory epithelial cells in sputum
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DOI：
10.1002/cyto.a.20041
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发表时间：
2004-07-01
期刊：
影响因子：
3.7
通讯作者：
Reid, BJ
中科院分区：
文献类型：
--
作者：
Kraemer, PS;Sanchez, CA;Reid, BJ
背景：与支气管镜活检和灌洗不同，诱导痰是一种容易获得的生物标本来源。然而，获得异常脱落细胞进行详细的分子研究是有限的，因为呼吸道上皮细胞仅占痰细胞群的约1%。我们开发了一种多参数流式分选策略来从非上皮痰细胞中纯化上皮细胞，使用抗细胞角蛋白抗体AE 1/AE 3来识别人上皮细胞和DAPI来染色DNA。我们排除了具有高度侧向散射的细胞，其主要由鳞状细胞和污染性巨噬细胞组成。剩余的细胞角蛋白阳性的呼吸道上皮细胞，然后根据抗细胞角蛋白（PE）与DNA（DAPI）parameters.Results：在这个证明的原则研究，AE 1AE 3细胞角蛋白/DNA流分选策略富集罕见的二倍体呼吸道上皮细胞从平均1.1%的细胞在未分选诱导痰样品的平均纯度为42%。因此，AE 1AE 3流式分选结果在38倍富集这些cells.Conclusions：我们报告了一种多参数流式细胞术检测和富集罕见的呼吸道上皮细胞诱导痰样品的平均纯度为42%。随着进一步的发展，这种方法可能是有用的一部分，在肺癌高危人群的分子筛选方法。(C)2004 Wiley-Liss，Inc.
Background: induced sputum, in contrast to bronchoscopic biopsies and lavages, is an easily obtained source of biological specimens. However, obtaining abnormal exfoliated cells for detailed molecular studies is limited because respiratory epithelial cells comprise only about 1% of sputum cell populations.Methods:. We developed a multiparameter flow sorting strategy to purify epithelial cells from nonepithelial sputum cells, using anti-cytokeratin antibody AE1/AE3 to recognize human epithelial cells and DAPI to stain DNA. We excluded cells with a high degree of side-scatter, which were composed predominantly of squamous cells and contaminating macrophages. The remaining cytokeratin-positive respiratory epithelial cells were then sorted based on anti-cytokeratin (PE) vs DNA (DAPI) parameters.Results: In this proof of principle study, the AE1AE3 cytokeratin/DNA flow sorting strategy enriched rare diploid respiratory epithelial cells from an average of 1.1% of cells in unsorted induced sputum samples to average purities of 42%. Thus, AE1AE3 flow-sorting results in a 38-fold enrichment of these cells.Conclusions: We report a multiparameter flow cytometric assay to detect and enrich rare respiratory epithelial cells from induced sputum samples to average purities of 42%. With further development, this methodology may be useful as part of a molecular screening approach of populations at high risk for lung cancer. (C) 2004 Wiley-Liss, Inc.