An integrated microfluidics platform with high-throughput single-cell cloning array and concentration gradient...

Area of Science:

• Biotechnology and Biomedical Engineering
• Cancer Research
• Pharmacology

Background:

• Tumor cell heterogeneity contributes to drug resistance, hindering effective cancer treatment.
• Current methods for assessing drug cytotoxicity at the single-cell level are limited in throughput and dynamic characterization.
• High-throughput single-cell analysis is crucial for developing precision cancer therapies.

Purpose of the Study:

• To develop a microfluidics-based platform for high-throughput, dynamic single-cell drug response analysis.
• To investigate the efficacy of drug combinations and single-agent treatments on leukemia cells.
• To characterize drug response heterogeneity in leukemia patients' primary cells.

Main Methods:

• Development of the "SMART" (Simple, Massive, Alive, Retainable, Trackable) microfluidics platform.
• Integration of a 4320-unit Microfluidic chamber Array (MAC) with a six-concentration gradient generator.
• Treatment of over 3000 single cells or cell clones with multiplex drug concentrations and dynamic gradient analysis.

Main Results:

• The SMART platform enables efficient analysis of drug effects on single leukemia cells and clones.
• Combined treatment with Imatinib and Resveratrol showed significantly higher efficacy (lower IC50) than single-agent treatments.
• Single-cell derived clones exhibited higher drug resistance (higher IC50) than individual single cells; patient-derived leukemia cells showed heterogeneity.

Conclusions:

• The microfluidics-based SMART platform facilitates high-throughput single-cell capture, culture, and dynamic drug response monitoring.
• This platform offers a novel approach for efficient investigation of anticancer drug effects.
• The technology holds promise for advancing drug discovery in leukemia and other cancers.