TY - JOUR
T1 - Classification of large circulating tumor cells isolated with ultra-high throughput microfluidic Vortex technology
AU - Che, James
AU - Yu, Victor
AU - Dhar, Manjima
AU - Renier, Corinne
AU - Matsumoto, Melissa
AU - Heirich, Kyra
AU - Garon, Edward B.
AU - Goldman, Jonathan
AU - Rao, Jianyu
AU - Sledge, George W.
AU - Pegram, Mark D.
AU - Sheth, Shruti
AU - Jeffrey, Stefanie S.
AU - Kulkarni, Rajan P.
AU - Sollier, Elodie
AU - Di Carlo, Dino
N1 - Funding Information:
The authors were supported by funding from the Office of Naval Research Young Investigator Program (#N000141210847), NIH IMAT Program (#5R33CA177456), the NIH IMAT Program, the Komen Foundation, and a sponsored research grant from Vortex Biosciences.
PY - 2016/3/15
Y1 - 2016/3/15
N2 - Circulating tumor cells (CTCs) are emerging as rare but clinically significant non-invasive cellular biomarkers for cancer patient prognosis, treatment selection, and treatment monitoring. Current CTC isolation approaches, such as immunoaffinity, filtration, or size-based techniques, are often limited by throughput, purity, large output volumes, or inability to obtain viable cells for downstream analysis. For all technologies, traditional immunofluorescent staining alone has been employed to distinguish and confirm the presence of isolated CTCs among contaminating blood cells, although cells isolated by size may express vastly different phenotypes. Consequently, CTC definitions have been non-trivial, researcher-dependent, and evolving. Here we describe a complete set of objective criteria, leveraging well-established cytomorphological features of malignancy, by which we identify large CTCs. We apply the criteria to CTCs enriched from stage IV lung and breast cancer patient blood samples using the High Throughput Vortex Chip (Vortex HT), an improved microfluidic technology for the label-free, size-based enrichment and concentration of rare cells. We achieve improved capture efficiency (up to 83%), high speed of processing (8 mL/min of 10x diluted blood, or 800 μL/min of whole blood), and high purity (avg. background of 28.8±23.6 white blood cells per mL of whole blood). We show markedly improved performance of CTC capture (84% positive test rate) in comparison to previous Vortex designs and the current FDA-approved gold standard CellSearch assay. The results demonstrate the ability to quickly collect viable and pure populations of abnormal large circulating cells unbiased by molecular characteristics, which helps uncover further heterogeneity in these cells.
AB - Circulating tumor cells (CTCs) are emerging as rare but clinically significant non-invasive cellular biomarkers for cancer patient prognosis, treatment selection, and treatment monitoring. Current CTC isolation approaches, such as immunoaffinity, filtration, or size-based techniques, are often limited by throughput, purity, large output volumes, or inability to obtain viable cells for downstream analysis. For all technologies, traditional immunofluorescent staining alone has been employed to distinguish and confirm the presence of isolated CTCs among contaminating blood cells, although cells isolated by size may express vastly different phenotypes. Consequently, CTC definitions have been non-trivial, researcher-dependent, and evolving. Here we describe a complete set of objective criteria, leveraging well-established cytomorphological features of malignancy, by which we identify large CTCs. We apply the criteria to CTCs enriched from stage IV lung and breast cancer patient blood samples using the High Throughput Vortex Chip (Vortex HT), an improved microfluidic technology for the label-free, size-based enrichment and concentration of rare cells. We achieve improved capture efficiency (up to 83%), high speed of processing (8 mL/min of 10x diluted blood, or 800 μL/min of whole blood), and high purity (avg. background of 28.8±23.6 white blood cells per mL of whole blood). We show markedly improved performance of CTC capture (84% positive test rate) in comparison to previous Vortex designs and the current FDA-approved gold standard CellSearch assay. The results demonstrate the ability to quickly collect viable and pure populations of abnormal large circulating cells unbiased by molecular characteristics, which helps uncover further heterogeneity in these cells.
KW - Circulating tumor cells
KW - Immunofluorescent staining
KW - Rare cell enrichment
KW - Size based cell isolation
KW - Vortex
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U2 - 10.18632/oncotarget.7220
DO - 10.18632/oncotarget.7220
M3 - Article
C2 - 26863573
AN - SCOPUS:84962895530
SN - 1949-2553
VL - 7
SP - 12748
EP - 12760
JO - Oncotarget
JF - Oncotarget
IS - 11
ER -