A Microfluidic-Biomaterial Platform for Personalized Diagnostics via Viable Circulating Tumor Cell Analysis in Colorectal Cancer

Colorectal cancer (CRC) remains one of the most lethal cancers globally, with increasing concern around delays in detection [1]. Circulating tumor cells (CTCs) offer a promising route for real-time, minimally invasive monitoring of cancer progression [2]. However, technical barriers related to their rarity and fragility limit current detection platforms [3]. In this study, we introduce a microfluidic device—ONCO-CTC tool—incorporating a removable electrospun polycaprolactone (PCL) scaffold designed to enhance CTC capture and post-isolation analysis.

The chip features a multilayer PDMS configuration and a fibrous insert engineered for optimal porosity and interconnectivity, promoting efficient cell retention and viability [4]. In vitro validation using HCT-116 CRC cells confirmed >94% capture efficiency and robust metabolic activity following 72 h culture. Compared to a commercial filtration system, the ONCO-CTC tool demonstrated superior performance in both recovery and cell preservation.

To assess clinical utility, blood samples from patients with metastatic CRC were processed through three protocols: RosetteSep™-based enrichment (control), conventional mononuclear (MNC) isolation, and ONCO-CTC-enabled capture. Cells identified as EpCAM+/CD45–/DAPI+ were detected across ONCO-CTC fractions, with high specificity (Figure 1). Importantly, captured cells were successfully cultured on membranes, and Cytospin analysis revealed maintained morphological integrity and marker expression for cytological analysis.This approach merges microengineering with functional biomaterials to achieve a user-friendly and modular tool for liquid biopsy applications. The ONCO-CTC tool supports both immediate CTC detection and downstream cellular analysis, reinforcing its promise as a personalized diagnostic aid for colorectal cancer management.

References:

[1] van den Puttelaar R. et al, 2023, 10.1158/1055-9965.EPI-22-0544

[2] Gu X. et al, 2024, 10.1038/s41392-024-01938-6

[3] Mishra A. et al, 2025, 10.1038/s41467-024-55140-x

[4] Casanova M.R. et al, 2024, Patent application No. 119865

Acknowledgments:

This research acknowledges the funding provided by UID/50026 (3B’s Research Group, University of Minho), CAN-TARGET (NORTE2030-FEDER-02705300, CCDR-N), and the European Commission through ONCOSCREEN (ID: 101097036) and EngVIPO (ID: 101183041) projects.