Isolation of circulating tumor cells by immunomagnetic enrichment and fluorescence-activated cell sorting (IE/FACS) for molecular profiling

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• 作者：Mark Jesus M. Magbanua ; John W. Park
• 关键词：Circulating tumor cells ; Fluorescence-activated cell sorting ; Immunomagnetic enrichment ; Array comparative genomic hybridization ; Single cell analysis
• 刊名：Methods
• 出版年：1 December, 2013
• 年：2013
• 卷：64
• 期：2
• 页码：114-118
• 全文大小：1401 K

文摘

Circulating tumor cells (CTCs) are cells shed by the primary tumor into the blood stream capable of initiating distant metastasis. In the past decade, numerous assays have been developed to reliably detect these extremely rare cells. However, methods for purification of CTCs with little or no contamination of normal blood cells for molecular profiling are limited. We have developed a novel protocol to isolate CTCs by combining immunomagnetic enrichment and fluorescence-activated cell sorting (IE/FACS). The two-part assay includes (1) immunomagnetic capture using magnetic beads conjugated to monoclonal antibody against an epithelial cell adhesion marker (EpCAM) to enrich for tumor cells; and (2) FACS analysis using EpCAM to purify tumor cells away from mononuclear cells of hematopoietic lineage. Downstream molecular analyses of single and pooled cells confirmed the isolation of highly pure CTCs with characteristics typical that of malignant cells.