1. Opportunities and Challenges for Pancreatic Circulating Tumor Cells
Sunitha Nagrath, Rhonda M. Jack, Vaibhav Sahai, Diane M. Simeone 
Gastroenterology 
Volume 151, Issue 3, Pages (September 2016)
DOI: /j.gastro
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2. Figure 1
Technologies used to enrich CTCs from patient samples. Immunoaffinity: (A) Positive selection, which can include immunomagnetic enrichment by targeting proteins expressed on CTCs; (B) negative selection eliminates nontarget cells such as leukocytes by use of antibodies against proteins expressed on cell surface such as CD45; (C) CTC immobilization involves cell adherence to antibody-coated surfaces based on proteins expressed on CTCs. Physical property: (D) Density gradient methods allow CTCs to separate out in media from other blood cells based on their densities. (E) Filtration uses a size-based filtering process to enrich CTCs from other blood cells based on their relatively larger radii compared with other cells. (F) Dielectrophoresis (DEP) utilizes the dielectric charge across cell membrane of CTCs unique from other cells in a conductivity medium. (G) Microfluidic hydrodynamics uses cell inertia to sort CTCs into distinct microfluidic channels from other blood cells.
Gastroenterology  , DOI: ( /j.gastro )
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3. Figure 2
Biophysical CTC characterization approaches include analysis of the following. (Ai) Physical dimensions such as radius, nuclear to cytoplasmic ratio, and sphericity. Adapted with permission from Lin et al.43 (Aii) Immunocytochemistry with H&E staining to reveal cell histology. Adapted with permission from Stott et al.35 (Aiii) Immunofluorescence typically for nuclear content (DAPI), cytokeratins, and negative control with the WBC marker CD45. Adapted with permission from Hou et al58 and Lim et al.134 (Aiv) RNA fluorescence in situ hybridization (FISH) to identify expression of RNA specific to proteins. Adapted with permission from Yu et al.89 Genomic characterization of CTCs includes the following. (Bi) RNA sequencing and expression profiles through the use of heat maps for CTC-enriched and EMT-specific transcripts. Adapted with permission from Yu et al.88 (Bii–iii) DNA sequencing of tumor and CTCs from the same patient to compare mutations such as deleted nucleotide sequences. Adapted with permission from Maheswaran et al.82 CTCs and clinical outcomes. (Ci) Evaluation of progression-free survival (PFS) of patients with pancreatic cancer with and without CTCs. Adapted with permission from de Albuquerque et al.100 (Cii) Evaluation of overall survival (OS) of patients undergoing chemotherapy as it relates to CTC status at baseline and/or 2 months: CTC positive (black line) versus CTC negative (gray line).
Adapted with permission from Bidard et al.27
Gastroenterology  , DOI: ( /j.gastro )
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