An ultra-high-throughput spiral microfluidic biochip for the enrichment of circulating tumor cells.
The detection and characterization of rare circulating tumor cells (CTCs) from the blood of cancer patients can potentially provide critical insights into tumor biology and hold great promise for cancer management. The ability to collect a large number of viable CTCs for various downstream assays such as quantitative measurements of specific biomarkers or targeted somatic mutation analysis is increasingly important in medical oncology. Here, we present a simple yet reliable microfluidic device for the ultra-high-throughput, label-free, size-based isolation of CTCs from clinically relevant blood volumes. The fast processing time of the technique (7.5 mL blood in less than 10 min) and the ability to collect more CTCs from larger blood volumes lends itself to a broad range of potential genomic and transcriptomic applications. A critical advantage of this protocol is the ability to return all fractions of blood (i.e., plasma (centrifugation), CTCs and white blood cells (WBCs) (size-based sorting)) that can be utilized for diverse biomarker studies or time-sensitive molecular assays such as RT-PCR. The clinical use of this biochip was demonstrated by detecting CTCs from 100% (10/10) of blood samples collected from patients with advanced-stage metastatic breast and lung cancers. The CTC recovery rate ranged from 20 to 135 CTCs mL(-1) and obtained under high purity (of 1 CTC out of every 30-100 WBCs which gives ∼4 log depletion of WBCs). They were identified with immunofluorescence assays (pan-cytokeratin+/CD45-) and molecular probes such as HER2/neu.
BioSystems and Micromechanics (BioSyM) IRG, Singapore-MIT Alliance for Research and Technology (SMART) Centre, Singapore.
Majid Ebrahimi
ME
Warkiani
Bee Luan
BL
Khoo
Daniel Shao-Weng
DS
Tan
Ali Asgar S
AA
Bhagat
Wan-Teck
WT
Lim
Yoon Sim
YS
Yap
Soo Chin
SC
Lee
Ross A
RA
Soo
Jongyoon
J
Han
Chwee Teck
CT
Lim
D001940:Breast / Q000473:pathology; D001943:Breast Neoplasms / Q000097:blood / Q000473:pathology; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000191:economics / Q000295:instrumentation*; D048429:Cell Size; D002470:Cell Survival; D004867:Equipment Design; D005260:Female; D006801:Humans; D008168:Lung / Q000473:pathology; D008175:Lung Neoplasms / Q000097:blood / Q000473:pathology; D046210:Microfluidic Analytical Techniques / Q000191:economics / Q000295:instrumentation*; D009362:Neoplasm Metastasis / Q000473:pathology; D009369:Neoplasms / Q000097:blood* / Q000473:pathology; D009360:Neoplastic Cells, Circulating / Q000473:pathology*