title: Classification of large circulating tumor cells isolated with ultra-high throughput microfluidic Vortex technology.
abstract: 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.
authors: Department of Bioengineering, University of California, Los Angeles, California, USA.|Vortex Biosciences, Menlo Park, California, USA.
authors: James
authors: J
authors: Che
authors: Department of Bioengineering, University of California, Los Angeles, California, USA.
authors: Victor
authors: V
authors: Yu
authors: Department of Bioengineering, University of California, Los Angeles, California, USA.
authors: Manjima
authors: M
authors: Dhar
authors: Vortex Biosciences, Menlo Park, California, USA.|Department of Surgery, Stanford University School of Medicine, Stanford, California, USA.
authors: Corinne
authors: C
authors: Renier
authors: Department of Bioengineering, University of California, Los Angeles, California, USA.
authors: Melissa
authors: M
authors: Matsumoto
authors: Department of Surgery, Stanford University School of Medicine, Stanford, California, USA.
authors: Kyra
authors: K
authors: Heirich
authors: Department of Hematology & Oncology, UCLA Medical Center, Los Angeles, California, USA.
authors: Edward B
authors: EB
authors: Garon
authors: Department of Hematology & Oncology, UCLA Medical Center, Los Angeles, California, USA.
authors: Jonathan
authors: J
authors: Goldman
authors: Department of Pathology & Laboratory Medicine, UCLA Medical Center, Los Angeles, California, USA.
authors: Jianyu
authors: J
authors: Rao
authors: Stanford Women's Cancer Center, Stanford, California, USA.
authors: George W
authors: GW
authors: Sledge
authors: Stanford Women's Cancer Center, Stanford, California, USA.
authors: Mark D
authors: MD
authors: Pegram
authors: Stanford Women's Cancer Center, Stanford, California, USA.
authors: Shruti
authors: S
authors: Sheth
authors: Department of Surgery, Stanford University School of Medicine, Stanford, California, USA.|Stanford Women's Cancer Center, Stanford, California, USA.
authors: Stefanie S
authors: SS
authors: Jeffrey
authors: Department of Bioengineering, University of California, Los Angeles, California, USA.|Division of Dermatology, UCLA Medical Center, Los Angeles, California, USA.
authors: Rajan P
authors: RP
authors: Kulkarni
authors: Department of Bioengineering, University of California, Los Angeles, California, USA.|Vortex Biosciences, Menlo Park, California, USA.|Department of Surgery, Stanford University School of Medicine, Stanford, California, USA.
authors: Elodie
authors: E
authors: Sollier
authors: Department of Bioengineering, University of California, Los Angeles, California, USA.|California NanoSystems Institue, Los Angeles, California, USA.|Jonsson Comprehensive Cancer Center, Los Angeles, California, USA.
authors: Dino
authors: D
authors: Di Carlo
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001943:Breast Neoplasms / Q000097:blood / Q000473:pathology; D005260:Female; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D008175:Lung Neoplasms / Q000097:blood / Q000473:pathology; D008297:Male; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D008875:Middle Aged; D009360:Neoplastic Cells, Circulating / Q000145:classification* / Q000473:pathology
keywords: Vortex; circulating tumor cells; immunofluorescent staining; rare cell enrichment; size based cell isolation