Microfluidic enrichment for the single cell analysis of circulating tumor cells

Abstract

Resistance to drug therapy is a major concern in cancer treatment. To probe clones resistant to chemotherapy, the current approach is to conduct pooled cell analysis. However, this can yield false negative outcomes, especially when we are analyzing a rare number of circulating tumor cells (CTCs) among an abundance of other cell types. Here, we develop a microfluidic device that is able to perform high throughput, selective picking and isolation of single CTC to 100% purity from a larger population of other cells. This microfluidic device can effectively separate the very rare CTCs from blood samples from as few as 1 in 20,000 white blood cells. We first demonstrate isolation of pure tumor cells from a mixed population and track variations of acquired T790M mutations before and after drug treatment using a model PC9 cell line. With clinical CTC samples, we then show that the isolated single CTCs are representative of dominant EGFR mutations such as T790M and L858R found in the primary tumor. With this single cell recovery device, we can potentially implement personalized treatment not only through detecting genetic aberrations at the single cell level, but also through tracking such changes during an anticancer therapy.

Biology and clinical significance of circulating tumor cell subpopulations in lung cancer

“…In a pilot study a spiral microfluidic device allowing high throughput, selective picking and isolation of single CTC was used to detect T790M mutations in seven NSCLC patients. This method also showed concordant results with the biopsy sample for six samples (48). The presence of the EGFR point mutation, T790M, in CTCs isolated from 27 NSCLC patients by CTC-chip was shown in another study to coincide with disease progression and reduced response to targeted therapies (17).…”

Clinical Utility of Circulating Tumor Cells – A Clinician’s Current View

“…In the recent literature from 2010-2015, over 35 different CTC methods were identified [4]. More new devices have been recently described [5][6][7][8][9]. CTC detection modalities have been well documented [10][11][12] and will not be covered in this review.…”

A Numerical Approach for Dielectrophoretic Characterization and Separation of Human Hematopoietic Cells

“…Characterization of cells is one of its applications, relies on its capability of generating specific dielectrophoretic forces ( F DEP ) for different types of cells, in (1).…”

“…Thanks to the developments in the lab-on-a-chip technologies, novel diagnostic methods intended to quantitatively characterize specific cell types in a fast and efficient way, directly from blood [1]. Integration of microfluidics with Micro-ElectroMechanical Systems (MEMS) provided portable, low-cost, high-throughput, fast and precise tools for medical diagnostics and biological sciences [2].…”