High-Viability Circulating Tumor Cells Sorting From Whole Blood at Single Cell Level Using Laser-Induced Forward...

Area of Science:

• Biomedical Engineering
• Cell Biology
• Oncology

Background:

• Efficient isolation of circulating tumor cells (CTCs) is vital for cancer metastasis research and personalized medicine.
• Cell viability is critical for downstream molecular analyses like RNA sequencing.
• Existing methods face challenges in achieving high-viability CTC isolation at single-cell resolution.

Purpose of the Study:

• To develop and validate a novel system for high-viability CTC enrichment and retrieval from whole blood.
• To enable downstream molecular analyses, including RNA sequencing, on isolated single CTCs.
• To improve the efficiency and throughput of CTC isolation for clinical applications.

Main Methods:

• Development of a laser-induced forward transfer-assisted microfiltration system (LIFT-AMFS).
• Utilized a LIFT-compatible double-stepped microfilter (DSMF) for size-based cell separation and liquid encapsulation.
• Optimized DSMF design for enhanced capture efficiency and single-cell retrieval.

Main Results:

• Achieved 88% CTC capture efficiency at a throughput of 15.0 mL/min.
• Obtained over 95% single-cell yield during the retrieval stage with high cell viability.
• Isolated CTCs yielded sufficient cDNA (>4.5 ng) for library construction, with Q30 scores >95.92% for single-cell sequencing.

Conclusions:

• The LIFT-AMFS effectively enriches and retrieves high-viability CTCs from whole blood.
• The system supports downstream molecular analyses, including ex vivo culture and RNA sequencing.
• LIFT-AMFS demonstrates significant potential for advancing cellular and biomedical research in oncology.