Classification of large circulating tumor cells isolated with ultra-high throughput microfluidic Vortex technology
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James Che1,2, Victor Yu1, Manjima Dhar1, Corinne Renier2,3, Melissa Matsumoto1, Kyra Heirich3, Edward B. Garon4, Jonathan Goldman4, Jianyu Rao5, George W. Sledge6, Mark D. Pegram6, Shruti Sheth6, Stefanie S. Jeffrey3,6, Rajan P. Kulkarni1,7, Elodie Sollier1,2,3 and Dino Di Carlo1,8,9
1 Department of Bioengineering, University of California, Los Angeles, California, USA
2 Vortex Biosciences, Menlo Park, California, USA
3 Department of Surgery, Stanford University School of Medicine, Stanford, California, USA
4 Department of Hematology & Oncology, UCLA Medical Center, Los Angeles, California, USA
5 Department of Pathology & Laboratory Medicine, UCLA Medical Center, Los Angeles, California, USA
6 Stanford Women’s Cancer Center, Stanford, California, USA
7 Division of Dermatology, UCLA Medical Center, Los Angeles, California, USA
8 California NanoSystems Institue, Los Angeles, California, USA
9 Jonsson Comprehensive Cancer Center, Los Angeles, California, USA
Dino Di Carlo, email:
Keywords: circulating tumor cells, immunofluorescent staining, rare cell enrichment, size based cell isolation, Vortex
Received: January 18, 2016 Accepted: January 21, 2016 Published: February 06, 2016
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.
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