Fast and efficient microfluidic cell filter for isolation of circulating tumor cells from unprocessed whole blood of colorectal cancer patients

Abstract

Liquid biopsy offers unique opportunities for low invasive diagnosis, real-time patient monitoring and treatment selection. The phenotypic and molecular profile of circulating tumor cells (CTCs) can provide key information about the biology of tumor cells, contributing to personalized therapy. CTC isolation is still challenging, mainly due to their heterogeneity and rarity. To overcome this limitation, a microfluidic chip for label-free isolation of CTCs from peripheral blood was developed. This device, the CROSS chip, captures CTCs based on their size and deformability with an efficiency of 70%. Using 2 chips, 7.5 ml of whole blood are processed in 47 minutes with high purity, as compared to similar technologies and assessed by in situ immunofluorescence. The CROSS chip performance was compared to the CellSearch system in a set of metastatic colorectal cancer patients, resulting in higher capture of DAPI+/CK+/CD45− CTCs in all individuals tested. Importantly, CTC enumeration by CROSS chip enabled stratification of patients with different prognosis. Lastly, cells isolated in the CROSS chip were lysed and further subjected to molecular characterization by droplet digital PCR, which revealed a mutation in the APC gene for most patient samples analyzed, confirming their colorectal origin and the versatility of the technology for downstream applications.

Evaluation of Microfluidic Ceiling Designs for the Capture of Circulating Tumor Cells on a Microarray Platform

“…These could potentially be used for cell culture and further downstream genomic analysis. Microfluidic systems have been shown to outperform the “gold standard” CellSearch device on multiple occasions . However, it is important to keep in mind, that most of these platforms utilize additional or alternative markers for CTC isolation or work with antibody cocktails, thereby complicating the direct comparison of both approaches.…”

Discrimination between Breast Cancer Cells and White Blood Cells by Non-Invasive Measurements: Implications for a Novel In Vitro-Based Circulating Tumor Cell Model Using Digital Holographic Cytometry

“…CTCs are usually larger in size, in contrast to leukocytes, and therefore, many CTCs isolation methods are mainly based on size differences [14,33,34]. A microcavity array (MCA) system based on a polycarbonate filter for the isolation by size of epithelial tumor cells has been developed [35,36]. The MCA system showed a significantly higher degree of CTCs isolation in advanced lung cancer patients than the CellSearch system [35].…”

New Insights Into the Role of Phenotypic Plasticity and EMT in Driving Cancer Progression

“…The FDA-approved CELLSEARCH R system (Menarini-Silicon Biosystems, Inc.), which immunocaptures EpCAM-expressing CTCs for patient prognosis (Riethdorf et al, 2007), is intrinsically biased toward predominantly epithelial CTCs. However, recent CTC studies have employed microfluidic devices to capture and isolate CTCs according to their size and deformability, which allows for better coverage of different phenotypic states (Lemaire et al, 2018;Ribeiro-Samy et al, 2019).…”