Design and Clinical Application of an Integrated Microfluidic Device for Circulating Tumor Cells Isolation and Single-Cell Analysis.

Mingxin Xu,Xinri Zhang,Kun Zou,Wenwen Liu,Song Wei,Qi Wang,Encheng Li

doi:10.3390/mi12010049

Abstract

Circulating tumor cells (CTCs) have been considered as an alternative to tissue biopsy for providing both germline-specific and tumor-derived genetic variations. Single-cell analysis of CTCs enables in-depth investigation of tumor heterogeneity and individualized clinical assessment. However, common CTC enrichment techniques generally have limitations of low throughput and cell damage. Herein, based on micropore-arrayed filtration membrane and microfluidic chip, we established an integrated CTC isolation platform with high-throughput, high-efficiency, and less cell damage. We observed a capture rate of around 85% and a purity of 60.4% by spiking tumor cells (PC-9) into healthy blood samples. Detection of CTCs from lung cancer patients demonstrated a positive detectable rate of 87.5%. Additionally, single CTCs, ctDNA and liver biopsy tissue of a representative advanced lung cancer patient were collected and sequenced, which revealed comprehensive genetic information of CTCs while reflected the differences in genetic profiles between different biological samples. This work provides a promising tool for CTCs isolation and further analysis at single-cell resolution with potential clinical value.

Highlights

Cancer is a major medical problem endangering human health, while metastasis remains the most common cause of tumor-related death
We report an integrated device based on micropore-arrayed filtration membrane and microfluidic chip to isolate circulating tumor cells (CTCs) for single-cell analysis
Since the size distributions of CTCs (12–25 μm) and white blood cells (WBCs) (5–20 μm) partially overlap [20], CTCs and some WBCs with larger size would be captured on the membrane

Summary

Introduction

Cancer is a major medical problem endangering human health, while metastasis remains the most common cause of tumor-related death. According to the “seed and soil” hypothesis proposed by Paget in 1889 [1], metastasis is a complex process when rare metastatic “seeds” shed from primary/secondary tumor lesions into the blood circulation and seek suitable “soil” to colonize. These “seeds”, called circulating tumor cells (CTCs), are considered as culprits of metastasis but a promising alternative to tissue biopsy [2]. For drug-related research, with microfluidics, a new approach has been developed and demonstrated for studying the physiological reactions of viable

Methods

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Conclusion