title: Adaptation of semiautomated circulating tumor cell (CTC) assays for clinical and preclinical research applications.
abstract: The majority of cancer-related deaths occur subsequent to the development of metastatic disease. This highly lethal disease stage is associated with the presence of circulating tumor cells (CTCs). These rare cells have been demonstrated to be of clinical significance in metastatic breast, prostate, and colorectal cancers. The current gold standard in clinical CTC detection and enumeration is the FDA-cleared CellSearch system (CSS). This manuscript outlines the standard protocol utilized by this platform as well as two additional adapted protocols that describe the detailed process of user-defined marker optimization for protein characterization of patient CTCs and a comparable protocol for CTC capture in very low volumes of blood, using standard CSS reagents, for studying in vivo preclinical mouse models of metastasis. In addition, differences in CTC quality between healthy donor blood spiked with cells from tissue culture versus patient blood samples are highlighted. Finally, several commonly discrepant items that can lead to CTC misclassification errors are outlined. Taken together, these protocols will provide a useful resource for users of this platform interested in preclinical and clinical research pertaining to metastasis and CTCs.
authors.affiliation: London Regional Cancer Program, London Health Sciences Centre; Department of Anatomy & Cell Biology, Schulich School of Medicine and Dentistry, Western University.
authors.forename: Lori E
authors.initials: LE
authors.lastname: Lowes
authors.affiliation: Special Hematology/Flow Cytometry, London Health Sciences Centre.
authors.forename: Benjamin D
authors.initials: BD
authors.lastname: Hedley
authors.affiliation: Special Hematology/Flow Cytometry, London Health Sciences Centre; Lawson Health Research Institute.
authors.forename: Michael
authors.initials: M
authors.lastname: Keeney
authors.affiliation: London Regional Cancer Program, London Health Sciences Centre; Department of Anatomy & Cell Biology, Schulich School of Medicine and Dentistry, Western University; Lawson Health Research Institute; Department of Oncology, Western University; alison.allan@lhsc.on.ca.
authors.forename: Alison L
authors.initials: AL
authors.lastname: Allan
mesh_terms: D000818:Animals; D001331:Automation; D003581:Cytodiagnosis / Q000295:instrumentation* / Q000379:methods*; D004195:Disease Models, Animal; D051379:Mice; D009362:Neoplasm Metastasis; D009374:Neoplasms, Experimental / Q000097:blood / Q000145:classification* / Q000473:pathology; D009360:Neoplastic Cells, Circulating / Q000145:classification* / Q000473:pathology*