Microfluidic Chip-Based Automatic System for Sequencing Patient-Derived Organoids at the Single-Cell Level.

Area of Science:

• Biotechnology
• Genomics
• Cancer Research

Background:

• Single-cell sequencing of patient-derived organoids (PDOs) is crucial for understanding organ heterogeneity and advancing cancer precision medicine.
• Existing methods for PDO single-cell sequencing are inefficient due to limited sample quantity and manual processes, hindering clinical application.

Purpose of the Study:

• To develop an automated microfluidic system for efficient and high-quality single-cell sequencing of PDOs.
• To address the limitations of current manual and microfluidic-based single-cell sequencing technologies for clinical tissue samples.

Main Methods:

• Development of a microfluidic chip-based automatic system named MASSO.
• Integration of PDO establishment, culturing, digesting, single-cell isolation, lysis, and whole-genome amplification within a single microfluidic chip.
• Characterization of the organoid single-cell whole-genome amplification chip (OSA-Chip) and the MASSO system.

Main Results:

• MASSO successfully performed whole-genome sequencing of lung cancer PDOs at the single-cell level, a novel achievement.
• The system avoids precious PDO loss and ensures high-quality on-chip whole-genome amplification.
• Automation by MASSO eliminated human error, improving data repeatability and bridging the gap between lab research and clinical practice.

Conclusions:

• MASSO provides a robust platform for single-cell whole-genome sequencing of PDOs, enabling precise identification of cancer-related and drug-response mutations.
• The system facilitates a deeper understanding of cell-level heterogeneities in PDOs and their original organs.
• MASSO lays the technical foundation for improved cancer precision medicine through efficient PDO analysis.