Enrichment and Detection of Circulating Tumor Cells and Other Rare Cell Populations by Microfluidic Filtration
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The current standard methods for isolating circulating tumor cells (CTCs) from blood involve EPCAM-based immunomagnetic approaches. A major disadvantage of these strategies is that CTCs with low EPCAM expression will be missed. Isolation by size using filter membranes circumvents the reliance on this cell surface marker, and can facilitate the capture not only of EPCAM-negative CTCs but other rare cells as well. These cells that are trapped on the filter membrane can be characterized by immunocytochemistry (ICC) , enumerated and profiled to elucidate their clinical significance. In this chapter, we discuss advances in filtration systems to capture rare cells as well as downstream ICC methods to detect and identify these cells. We highlight our recent clinical study demonstrating the feasibility of using a novel method consisting of automated microfluidic filtration and sequential ICC for detection and enumeration of CTCs, as well as circulating mesenchymal cells (CMCs), circulating endothelial cells (CECs), and putative circulating stem cells (CSCs). We hypothesize that simultaneous analysis of circulating rare cells in blood of cancer patients may lead to a better understanding of disease progression and development of resistance to therapy.
KeywordsCirculating tumor cells Circulating mesenchymal cells Circulating endothelial cells Circulating stem cells Filtration Immunocytochemistry Filter membranes
We acknowledge the research contributions by colleagues at Siemens Healthcare Diagnostics, namely, Mary Foltz, Karen Marfurt, Cathleen Hanau-Taylor, Julia Philps. We thank Jin Sun Lee, Marc Jabon, Victoria Wang, Matthew Gubens, and Hope S. Rugo at the University of California San Francisco for their assistance in the clinical work. MJM was supported in part by the Breast Cancer Research Foundation.